Difference between revisions of "Template:NTHU Taiwan/js/bundle"

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/*! *****************************************************************************
 
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and limitations under the License.
 
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object-assign
 
object-assign
Line 25: Line 25:
 
@license MIT
 
@license MIT
 
*/
 
*/
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Therefore, our goal this year is to design a system which is able to dose appropriate and corrected aquacultural drugs according to the concentration of bacteria in the water as well as automatically detect the water condition which can let fishermen have a comprehensive understanding of whether the drug works or not.")),c=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},"Our project can be divided into two parts: one is to dose appropriate and corrected drug, and the other is to automatically detect the water quality and the concentration of specific bacteria. "),a.default.createElement("div",{className:"part"},"To implement the idea of appropriately dosing aquaculture drug, we design a system based on the model of succession. 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The system regulates the rate of killing ",a.default.createElement("span",{className:"special-name"},"Vibrio")," and ",a.default.createElement("span",{className:"special-name"},"E. coli")," itself in order to keep the population of ",a.default.createElement("span",{className:"special-name"},"Vibrio")," under the pathogenic level and maintain the balance between two bacteria at the same time. Therefore, we can control the ",a.default.createElement("span",{className:"special-name"},"Vibrio")," population and avoid fish and shrimp infection in a sustainable way. "),a.default.createElement("div",{className:"part"},"In the part of detecting water quality, to get precise concentration of specific bacteria in the water, we design an automatic and real-time detecting device. The current detection methods are time-consuming and furthermore, the water quality monitoring system on the market currently is able to detect physical and chemical indexes only. Therefore, we aim to create an IoT detecting device which can collect the water sample automatically and periodically. Then it is able to detect the concentration of specific bacteria with high specificity. For instance, it can detect the concentration of 100 CFU/mL of ",a.default.createElement("span",{className:"special-name"},"Vibrio")," within 24 hours[4,5], and informs fisherman regularly. It has the potential to reduce the financial loss of our aquaculture industry. ")),f=a.default.createElement("div",null,a.default.createElement("div",{className:"reference"},"1. Ching-Ying Deng, Meei-Mei Chen, Ju-Hsien Peng, Yu-Chiao Yang, Chih-Chien Chang, Hui-Ping Tsai 2014 The Investigation on the Infection of Francisellosis in Farmed Tilapia in Taoyuan and Hsinchu Areas. Journal of Taiwan Fisheries Research,22 (1): 81-91"),a.default.createElement("div",{className:"reference"},"2. https://news.un.org/en/audio/2014/05/588932, UN NEWS. 19 May 2014 "),a.default.createElement("div",{className:"reference"},"3. H-J Ju, M-Y Huang, H-C Liu, L-W Tseng, C-L Pan and C-I Chang, 2016 Screening Bacteria Antagonistic toward Aquatic Pathogenic Vibrio spp. from Marine Fishes and Shrimp. Journal of Taiwan Fisheries Research, 24 (1): 37-50"),a.default.createElement("div",{className:"reference"},"4. Chin-I Chan, et al., 2011, https://www.tfrin.gov.tw/dl.asp?fileName=17111695971.pdf"),a.default.createElement("div",{className:"reference"},"5. http://www.tfrin.gov.tw/friweb/frienews/enews0118/h1.html")),d={viewbar:{Title:"Description",Subtitle:"A solution to the problem of the abuse of antibiotics in aquaculture",image:o.default.image.topfull.description},content:{Introduction:"",parts:[{Title:"Motivation",Description:u},{Title:"Solution",Description:c},{Title:"Reference",Description:f}]}},p=function(e){function t(){return function(e,t){if(!(e instanceof t))throw new TypeError("Cannot call a class as a function")}(this,t),function(e,t){if(!e)throw new ReferenceError("this hasn't been initialised - super() hasn't been called");return!t||"object"!=typeof t&&"function"!=typeof t?e:t}(this,(t.__proto__||Object.getPrototypeOf(t)).apply(this,arguments))}return function(e,t){if("function"!=typeof t&&null!==t)throw new TypeError("Super expression must either be null or a function, not "+typeof t);e.prototype=Object.create(t&&t.prototype,{constructor:{value:e,enumerable:!1,writable:!0,configurable:!0}}),t&&(Object.setPrototypeOf?Object.setPrototypeOf(e,t):e.__proto__=t)}(t,i.Component),r(t,[{key:"render",value:function(){return a.default.createElement(l.default,d)}}]),t}();t.default=p},function(e,t,n){"use strict";Object.defineProperty(t,"__esModule",{value:!0}),t.default=void 0;var r=function(){function e(e,t){for(var n=0;n<t.length;n++){var r=t[n];r.enumerable=r.enumerable||!1,r.configurable=!0,"value"in r&&(r.writable=!0),Object.defineProperty(e,r.key,r)}}return function(t,n,r){return n&&e(t.prototype,n),r&&e(t,r),t}}(),i=n(0),a=s(i),o=s(n(4)),l=s(n(8));function s(e){return e&&e.__esModule?e:{default:e}}var u=a.default.createElement("div",null,a.default.createElement("div",null,"Our project this year is to design an engineered E.coli to co-culture with pathogenic Vibrio and create a prey-predator system, also called bio-control, to control Vibrio population in order to create an equilibrium between Vibrio and engineered E.coli. Therefore Vibrio population can be controlled in a particular concentration which will not able to infect marine organism such as shrimp, fish…etc."),a.default.createElement("div",null,a.default.createElement("div",{className:"illustration"},a.default.createElement("div",{className:"illust-image-wrapper"},a.default.createElement("img",{className:"illust-image",src:o.default.image.design.exp})),a.default.createElement("div",{className:"undertext"},"Fig 1. exponential growth of vibrio without constraint in fish farm")),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.design.equlibrium}),a.default.createElement("div",{className:"undertext"},"Fig 2. Vibrio population controlled by engineered E.coli forming a equilibrium")),a.default.createElement("div",null)),a.default.createElement("div",{className:"illustration"},a.default.createElement("div",{className:"illust-image-wrapper"},a.default.createElement("img",{className:"illust-image",src:o.default.image.design.shrimp})),a.default.createElement("div",{className:"undertext"},"Fig 3. Infected shrimp")),a.default.createElement("div",null,"Hence the goal of wetlab this year is going to design a gene circuit which can make engineered E.coli to secrete anti-vibrio substances[1] while vibrio concentration exceed hazard concentration, and engineered E.coli suicide[2] while vibrio concentration is low to avoid bio-safety concern."),a.default.createElement("div",{className:"illustration"},a.default.createElement("div",{style:{margin:"auto",width:"500px"}},a.default.createElement("img",{className:"illust-image",src:o.default.image.design.cycle}))),a.default.createElement("div",{className:"part"}," Our gene design can be divided into 3 parts. First is the “detection” part makes our E.coli sense the population of vibrio and respond to it.  Second, the “kill switch” part compares the level of 2 bacteria and switches our E.coli  between modes of secreting anti-vibrio substances and suicide substance. “Growth inhibitor” part includes a toxic protein (so called Colicin M) and a toxic small RNA (tsRNA)."),a.default.createElement("div",{className:"part"}," Though we want to conduct Vibrio related experiments in our lab, our lab belongs to BSL1 level and we can’t cultivate Vibrio. Therefore, we use 2 different kinds of E. coli. to simulate our succession model with one being as fake Vibrio, and another being as engineered E. coli. ")),c=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},"ACE is an abbreviation of “Automatic Color-detecting Examiner” which is able to provide a stable environment for any color-changing-related test. All of the processes are executed and the information of each color will be digitized automatically. Finally, we create our own database to store the data and analyze them more precisely. Combining with IoT technology, all of our data will be recorded and analyzed in our cloud database and be returned to our clients, which allows them to take control of the results."),a.default.createElement("div",{className:"part"},"In order to prove our ACE can be used in reality to solve the Vibrio problem in our society, we make a collaboration with the “Fisheries Research Institute” of the Taiwan government. 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OD 600 Reference point"),a.default.createElement("div",null,"LUDOX CL-X (45% colloidal silica suspension) is used as a single point reference to obtain a conversion factor to transform absorbance (Abs600) data from our plate reader into a comparable OD600 measurement as would be obtained in a spectrophotometer. We will get the data for OD600 of the H2O and LUDOX. The corrected Abs600 is calculated by subtracting the H2O reading. To convert measured Abs600 to OD600 is to let Reference OD600 divided by Abs600.")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"2. Particle Standard Curve"),a.default.createElement("div",null,"We prepared a dilution series of monodisperse silica microspheres and measured the Abs600 in a plate reader. The size and optical characteristics of these microspheres are similar to cells, and there was a known amount of particles per volume. This measurement allows us to construct a standard curve of particle concentration which can be used to convert Abs600 measurements to an estimated number of cells.")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"3. Fluorescence standard curve"),a.default.createElement("div",null,"Absolute fluorescence values cannot be directly compared from one instrument to another. In order to compare the fluorescence output of test devices between teams, it was necessary to create a standard fluorescence curve.")))),f=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},a.default.createElement("div",null,"First, we transformed Escherichia coli DH5α with these following plasmids."),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",null,"Negative control  BBa_R0040    Kit Plate 7 Well 2D"),a.default.createElement("div",null,"Positive control  BBa_I20270  Kit Plate 7 Well 2B"),a.default.createElement("div",null,"Test Device 1  BBa_J364000  Kit Plate 7 Well 2F"),a.default.createElement("div",null,"Test Device 2  BBa_J364001  Kit Plate 7 Well 2H"),a.default.createElement("div",null,"Test Device 3  BBa_J364002  Kit Plate 7 Well 2J"),a.default.createElement("div",null,"Test Device 4  BBa_J364007  Kit Plate 7 Well 2L"),a.default.createElement("div",null,"Test Device 5      BBa_J364008  Kit Plate 7 Well 2N"),a.default.createElement("div",null,"Test Device 6      BBa_J364009  Kit Plate 7 Well 2P"))),a.default.createElement("div",{className:"part"},"Secondly, two colonies from each plate were picked and inoculated in LB medium containing chloramphenicol overnight for 16-18 hours at 37°C and 220 rpm."),a.default.createElement("div",{className:"part"},"Finally, we measured the OD600 and fluorescence of transformed cells according to the protocol after 0, 6 hours. 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Outside of iGEM, he is now joining in an immunology laboratory and doing some researches about immune checkpoints, PD-1. Apart from the university studies, he is also active in diverse activities. For example, he has participated in several Chinese calligraphy contests in Taiwan and won many prizes.",contact:{facebook:"https://www.facebook.com/tysh0106",linkedin:"https://www.linkedin.com/in/tysh0106/"}},{name:"Kevin Wang",image:""+C.default.image.member.Kevin_Wang,position:"Wetlab",department:"Materials Science and Engineering/ Physics",description:"I am Kevin Wang, a sophomore student study in Material Sciences and Physics. \n      During iGEM, I design the gene circuit and most of gene fragments of our project this year, and including trouble shooting. This summer, I spent lots of time doing digestion-ligation, transform and PCR. Although failure make me feel frustration, I never give up till success. 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Also, Opera and EDM are the two music genres she loves most : )",contact:{facebook:"https://www.facebook.com/tinawu2406",linkedin:"https://www.linkedin.com/in/xin-fang-wu/",email:"tinawu2406@gmail.com"}},{name:"Ruby Chan",image:""+C.default.image.member.Ruby_Chan,position:"Wetlab",department:"Chemistry",description:"A girl who is never satisfied with the status quo.",contact:{facebook:"https://www.facebook.com/profile.php?id=100004026627188"}},{name:"Andy Yu",image:""+C.default.image.member.Kai_Chiang_Yu,position:"Wetlab",department:"Chemistry",description:"Andy, a senior student from department of chemistry, now struggling along the tenebrous road from hell.",contact:{facebook:"https://www.facebook.com/profile.php?id=100002090493215"}},{name:"Ricky Chan",image:""+C.default.image.member.Ricky_Chan,position:"Wetlab",department:"Applied Science",description:"A boy who is curious about this unknown world, and likes to connect with different people. 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Combining with IoT technology, all of our data will be recorded and analyzed in our cloud database and be returned to our clients, which allows them to take control of the results."),o.default.createElement("div",{className:"illustration"},o.default.createElement("div",{className:"illust-image-wrapper"},o.default.createElement("img",{className:"illust-image",src:s.default.image.drylab_overview.box})))),h=o.default.createElement("div",null,o.default.createElement("div",{className:"part"},o.default.createElement("div",null,"In order to prove our ACE can be used in reality to solve the problems in our society, we make a collaboration with the “Fisheries Research Institute” of the Taiwan government. They kindly provided us with their “Multiple-Vibrio Detecting Kit, MV-kit” and we integrated their MV-kit with our system to develop a brand-new",o.default.createElement("a",{href:"https://2018.igem.org/Team:NTHU_Taiwan/Application"}," Automatic Vibrio-detecting System"),".")),o.default.createElement("div",{className:"part"},"Through the optimization of ACE, we accelerate the detection rate of MV-kit. With our ACE, it becomes 2 times faster than the original MV-kit and the minimal detectable Vibrio concentration is about 100 CFU/ mL within 24 hours. In summary, we combine the traditional detecting methods with the cutting-edge technology to achieve a better effect. In addition, following with our IoT system makes the water quality testing system in aquatic agriculture industry step forward to the whole new world of “intelligent aquaculture”."),o.default.createElement("div",{className:"illustration"},o.default.createElement("img",{className:"illust-image",src:s.default.image.drylab_overview.plate}))),m=o.default.createElement("div",null,o.default.createElement("div",null,"Components list and cost on ",o.default.createElement("a",{href:" https://docs.google.com/spreadsheets/d/1NsoRao-FtywW1KCeFYrxMIjJG3kVGQRcg_fIhxjn7ZQ/edit?usp=sharing"},"Google drive")),o.default.createElement("div",null,"3D printing files on ",o.default.createElement("a",{href:"https://drive.google.com/drive/folders/1tPQEVtNHB_hzhalPlwZu4bzyQVSDUPkN?usp=sharing"},"Google Drive"))),g={viewbar:{Title:"Drylab",Subtitle:"ACE- Automatic Color-detecting Examiner",image:s.default.image.topfull.drylab_overview,Underimage:f},content:{Introduction:d,parts:[{Title:"ACE",Description:p},{Title:"Application",Description:h},{Title:"File download",Description:m}]}},v=(0,l.inject)("appState")(r=(0,l.observer)(r=function(e){function t(e){return function(e,t){if(!(e instanceof t))throw new TypeError("Cannot call a class as a function")}(this,t),function(e,t){if(!e)throw new ReferenceError("this hasn't been initialised - super() hasn't been called");return!t||"object"!=typeof t&&"function"!=typeof t?e:t}(this,(t.__proto__||Object.getPrototypeOf(t)).call(this,e))}return function(e,t){if("function"!=typeof t&&null!==t)throw new TypeError("Super expression must either be null or a function, not "+typeof t);e.prototype=Object.create(t&&t.prototype,{constructor:{value:e,enumerable:!1,writable:!0,configurable:!0}}),t&&(Object.setPrototypeOf?Object.setPrototypeOf(e,t):e.__proto__=t)}(t,a.Component),i(t,[{key:"render",value:function(){return o.default.createElement(u.default,g)}}]),t}())||r)||r;t.default=v},function(e,t,n){"use strict";Object.defineProperty(t,"__esModule",{value:!0});var r=function(){function e(e,t){for(var n=0;n<t.length;n++){var r=t[n];r.enumerable=r.enumerable||!1,r.configurable=!0,"value"in r&&(r.writable=!0),Object.defineProperty(e,r.key,r)}}return function(t,n,r){return n&&e(t.prototype,n),r&&e(t,r),t}}(),i=n(0),a=s(i),o=(s(n(10)),s(n(4))),l=s(n(8));function s(e){return e&&e.__esModule?e:{default:e}}var u=a.default.createElement("div",null,a.default.createElement("div",null,"After extracting a sample from the fish farm, the sample will be dripped into the 96 -well plate. In the plate, we put in the mixture of MV-kit buffer for the sample to react. There are several reasons we chose to use 96-well plates instead of eppendorf or 6/24/48 well plate."),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"innerpart"},"1. Eppendorf has an unremovable lid, which is not convenient for our auto sampling design."),a.default.createElement("div",{className:"innerpart"},"2. The base of eppendorf is sharp and liquid will not distribute evenly, which is not suitable for color sensing while 96-well plates are flat."),a.default.createElement("div",{className:"innerpart"},"3. The volume of the 96-well plate is small enough for us to put the sample in, while the other plates are too big."),a.default.createElement("div",{className:"innerpart"},"4. It can contain the most samples in one single plate. In that way, we can reduce the rate of changing new plates.")),a.default.createElement("div",{className:"illustration"},a.default.createElement("table",{className:"table"},a.default.createElement("tbody",null,a.default.createElement("tr",null,a.default.createElement("th",{className:"tablehead"},"Number of well"),a.default.createElement("th",{className:"tablehead"},"Radius (mm) "),a.default.createElement("th",{className:"tablehead"},"Max Volume (mL) "),a.default.createElement("th",{className:"tablehead"},"Growing Area (cm2)")),a.default.createElement("tr",null,a.default.createElement("td",{className:"tabledata"},"96"),a.default.createElement("td",{className:"tabledata"},"4.5 "),a.default.createElement("td",{className:"tabledata"},"0.36"),a.default.createElement("td",{className:"tabledata"},"0.16")),a.default.createElement("tr",null,a.default.createElement("td",{className:"tabledata"},"48"),a.default.createElement("td",{className:"tabledata"},"10.2"),a.default.createElement("td",{className:"tabledata"},"1.7"),a.default.createElement("td",{className:"tabledata"},"0.8")),a.default.createElement("tr",null,a.default.createElement("td",{className:"tabledata"},"24"),a.default.createElement("td",{className:"tabledata"},"15.6"),a.default.createElement("td",{className:"tabledata"},"3.4"),a.default.createElement("td",{className:"tabledata"},"1.9")),a.default.createElement("tr",null,a.default.createElement("td",{className:"tabledata"},"12"),a.default.createElement("td",{className:"tabledata"},"22.1"),a.default.createElement("td",{className:"tabledata"},"6.9"),a.default.createElement("td",{className:"tabledata"},"3.8")),a.default.createElement("tr",null,a.default.createElement("td",{className:"tabledata"},"6"),a.default.createElement("td",{className:"tabledata"},"34.8"),a.default.createElement("td",{className:"tabledata"},"16.8"),a.default.createElement("td",{className:"tabledata"},"9.5")))),a.default.createElement("div",{className:"undertext"},"reference: http://www.dxy.cn/bbs/topic/9584997"),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.platform.transparent})),a.default.createElement("div",{className:"undertext"},"reference: http://www.carelifetech.com.tw/?96%E5%AD%94%E7%9B%A4-96-well-plate,127"))),c=a.default.createElement("div",null,a.default.createElement("div",null,"Since we need to use the camera to detect the color changes of each well, we need to create an environment that can make the color as clear as possible.  Therefore, we use 3D printing and polylactic acid (PLA) to print out a platform that blacks out all the area except for the 24 holes of each plate that have our sample in."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.platform.block}))),f=a.default.createElement("div",null,a.default.createElement("div",null,"Since the MV-kit buffer and solution might evaporate, we need to construct an auto moving lid. The lid has 48 holes on it, each hole is 9.1 mm next to the well. This lid is connected with a gear and a stepper motor. Every time the sample is being dripped into the well, the motor will move the lid by 9.1 mm. The gear system and the base of the motor are also made by 3D printing. Moreover, we’ve designed the lid to be detachable so that the farmer can remove the lid when they are going to substitute the 96 well plates with a new one every two weeks. "),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.platform.moving}))),d={viewbar:{Title:"Platform",Subtitle:"Plate and the moving lid",image:o.default.image.topfull.platform},content:{Introduction:"The reaction platform mainly consists of three parts: The 96-Well Microtitration Plate, the light blocking base, and auto moving lid.",parts:[{Title:"96-Well Microtitration",Description:u},{Title:"Light blocking",Description:c},{Title:"Moving lid",Description:f}]}},p=function(e){function t(){return function(e,t){if(!(e instanceof t))throw new TypeError("Cannot call a class as a function")}(this,t),function(e,t){if(!e)throw new ReferenceError("this hasn't been initialised - super() hasn't been called");return!t||"object"!=typeof t&&"function"!=typeof t?e:t}(this,(t.__proto__||Object.getPrototypeOf(t)).apply(this,arguments))}return function(e,t){if("function"!=typeof t&&null!==t)throw new TypeError("Super expression must either be null or a function, not "+typeof t);e.prototype=Object.create(t&&t.prototype,{constructor:{value:e,enumerable:!1,writable:!0,configurable:!0}}),t&&(Object.setPrototypeOf?Object.setPrototypeOf(e,t):e.__proto__=t)}(t,i.Component),r(t,[{key:"render",value:function(){return a.default.createElement(l.default,d)}}]),t}();t.default=p},function(e,t,n){"use strict";Object.defineProperty(t,"__esModule",{value:!0});var r=function(){function e(e,t){for(var n=0;n<t.length;n++){var r=t[n];r.enumerable=r.enumerable||!1,r.configurable=!0,"value"in r&&(r.writable=!0),Object.defineProperty(e,r.key,r)}}return function(t,n,r){return n&&e(t.prototype,n),r&&e(t,r),t}}(),i=n(0),a=s(i),o=(s(n(10)),s(n(8))),l=s(n(4));function s(e){return e&&e.__esModule?e:{default:e}}var u=a.default.createElement("div",null,"For the results of color-changing, we write a program to detect them. In our device, there are 24 samples processing the chemical reaction simultaneously. During the reaction, we want to get all samples’ color values immediately, so we adopt image detection executed by Raspberry Pi to analyze the whole status directly. The following are three parts for our image detection, which is how to do, advantages, and problems respectively."),c=a.default.createElement("div",null,a.default.createElement("div",null,"The following is the materials image detection needs"),a.default.createElement("table",{className:"table"},a.default.createElement("tbody",null,a.default.createElement("tr",null,a.default.createElement("td",{className:"tabledata"},"Raspberry Pi 3 Model B v1.3"),a.default.createElement("td",{className:"tabledata"},"1")),a.default.createElement("tr",null,a.default.createElement("td",{className:"tabledata"},"Webcam Logitech C270"),a.default.createElement("td",{className:"tabledata"},"1")),a.default.createElement("tr",null,a.default.createElement("td",{className:"tabledata"},"Wide-angle lens 37mm 0.6X"),a.default.createElement("td",{className:"tabledata"},"1")))),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:l.default.image.imaging.materials})),a.default.createElement("div",{className:"part"},"The core of image detection is executed by Raspberry Pi and the program language we use is python. With a camera and wide-angle lens, it can capture the scene in a relatively lower height. To do the image processing, OpenCV, open source library, is the best choice. Because the program will send warning message to the users, for simplicity and stability, we choose MQTT to be the transport protocol."),a.default.createElement("div",{className:"part"},"The following three steps are the processes our image detection works on. First, it captures the scene using a camera in a specified time. Second, the image is analyzed immediately after one is captured and the results are record. Third, after the analysis, if there is a color-changing sample, it will send the warning message to the users. All the steps are operated recursively until all the samples are analyzed.")),f=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},"For our image detection, we have three advantages, which are accuracy, automation, and flexibility, respectively. With these three, it can match our goal of designing ACE, i.e., automation and also increase usability compared to other devices used in the laboratory nowadays."),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"1. Accuracy"),a.default.createElement("div",{className:"innerpart"},"Because our 24 samples are close to each other, the accuracy is necessary to avoid the wrong result of analysis. With image detection, we can set the section where we want to analyze corresponding to the samples on the picture. Therefore, if all samples are located in advance carefully, we can get high accuracy for the results.")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"2. Automation"),a.default.createElement("div",{className:"innerpart"},"Automation is our goal of designing the device. To achieve this, the program can automatically do the following things, i.e., capturing the scene, analyzing the captured image, recording the results, and sending the warning messages to users. With the four steps as mentioned above operated cyclically, the program achieves automation.")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"3. Flexibility"),a.default.createElement("div",{className:"innerpart"},"The section program analyzes on the captured image can be modified whenever the captured one is changed. Therefore, we don’t have to fix the position of all samples. Due to this advantage, any type of sample can be analyzed accurately."))),d=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},"During the development of image detection, we had made some choices to solve many problems, including the specific practice of detection, development board, the algorithm to analyze the images, and the environment light. The following describes each problem, and the reasons why we made these choices."),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"1. Image detection vs. Color sensor"),a.default.createElement("div",{className:"innerpart"},"Initially, color sensor is our direction to design the device. Moving the color sensor to the top of each sample can get the color of that. However, our samples are close to each other, and the resolution of a color sensor is not high enough to detect the color of single sample accurately, so the direction is changed to image detection with the camera capturing the scene. In this way, it analyzes the image directly, the accuracy increases by a large margin.")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"2. Raspberry Pi vs. Other development boards"),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:l.default.image.imaging.board})),"For the development boards, there are three choices, Arduino, Udoo Neo, and Raspberry Pi. Considering the efficiency, Arduino might not be able to run OpenCV effectively. The another consideration is the cost. Because we want to design a low-cost device cheaper than the similar machine sold on the market, the price of Udoo Neo is too expensive for us. Therefore, we chose Raspberry Pi to be our execution core.")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"3. Simplified location vs. Houghcircles"),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:l.default.image.imaging.algorithm})),"In the beginning, we planned to use OpenCV built-in function Houghcircles to do image detection. The detail of the function is that it can find circles on the image automatically. It completely meets our expectation. However, the distance and angle of each sample from the camera is different. As a result, the shape of the sample is not a circle on the captured image and leads to wrong detection by Houghcircles. For this reason, we changed the method and decided to locate the samples manually to get a high accuracy."))),p=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:l.default.image.imaging.light})),"In order to prevent the influence of environment outside the device, ACE is designed to be enclosed. Therefore, it is completely dark inside the device and needs a light source to do image detection. In the beginning, we thought a big bulb is simple and suitable. Nevertheless, after we tested, we found the light on the image is not even, it would result in the wrong analysis. As a result, we chose LED strip. Its light is not only even but also bright enough.")),h=a.default.createElement("div",null,"You can download the source of our program on",a.default.createElement("a",{href:"https://github.com/igem-tw-nthu/opencv"}," Github")),m={viewbar:{Title:"Imaging",Subtitle:"Imaging by Raspberry Pi with OpenCV",image:l.default.image.topfull.imaging},content:{Introduction:u,parts:[{Title:"How to Do",Description:c},{Title:"Advantages",Description:f},{Title:"Problems",Description:d},{Title:"LED strip vs. Big bulb",Description:p},{Title:"Code Download",Description:h}]}},g=function(e){function t(){return function(e,t){if(!(e instanceof t))throw new TypeError("Cannot call a class as a function")}(this,t),function(e,t){if(!e)throw new ReferenceError("this hasn't been initialised - super() hasn't been called");return!t||"object"!=typeof t&&"function"!=typeof t?e:t}(this,(t.__proto__||Object.getPrototypeOf(t)).apply(this,arguments))}return function(e,t){if("function"!=typeof t&&null!==t)throw new TypeError("Super expression must either be null or a function, not "+typeof t);e.prototype=Object.create(t&&t.prototype,{constructor:{value:e,enumerable:!1,writable:!0,configurable:!0}}),t&&(Object.setPrototypeOf?Object.setPrototypeOf(e,t):e.__proto__=t)}(t,i.Component),r(t,[{key:"render",value:function(){return a.default.createElement(o.default,m)}}]),t}();t.default=g},function(e,t,n){"use strict";Object.defineProperty(t,"__esModule",{value:!0});var r=function(){function e(e,t){for(var n=0;n<t.length;n++){var r=t[n];r.enumerable=r.enumerable||!1,r.configurable=!0,"value"in r&&(r.writable=!0),Object.defineProperty(e,r.key,r)}}return function(t,n,r){return n&&e(t.prototype,n),r&&e(t,r),t}}(),i=n(0),a=u(i),o=(u(n(10)),u(n(4))),l=u(n(8)),s=u(n(136));function u(e){return e&&e.__esModule?e:{default:e}}var c=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},a.default.createElement("div",null,"For most of the detection reagent, they can only react with the small amount of sample each time. Taking MV-kit (Multiple Vibrio Detecting Kit) that we’re using for vibrio detection for example, it needs to have fix amount of 35μl sample fluid being collected every time so that we can get the better result of the later on reaction with the reagent. We choose mini peristaltic pump to fulfill our need. There are a few advantages of choosing peristaltic pump as our dispensing media[1]:"),a.default.createElement("div",null,a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",{className:"part-title"},"1. Accurate metering "),a.default.createElement("div",null,"Peristaltic pumps use flexible tubing to run through rollers in the pump head. As fluid goes through the pump head, packet of fluids can be formed. Simply by determining the size of the tubing and the speed of the roller turning in the pump head, the accurate metering of the media can be achieved.")),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",{className:"part-title"},"2. Self priming "),a.default.createElement("div",null,"Peristaltic pumps can draw fluid into the tubing when starting dry while other pumps require user to fill pump and suction line with fluid before use which can be inconvenient and also create potential for contamination.")),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",{className:"part-title"},"3. Non-siphoning "),a.default.createElement("div",null,"A peristaltic pump is non-siphoning which means that it prevents back flow into the system when the pump stop turning. This promotes accuracy during dispensing.")))),a.default.createElement("div",{className:"part"},a.default.createElement("div",null,"As a result, peristaltic pumps are often used for applications involving metering, dispensing and precise general transfer. Here, we use it in our device and we get the stl. files from the open source of the YouTube channels called “electronic crafts”[2]. The dimension of our peristaltic pump is 31 x 31 x 11 (mm), and the detail of other components are showed in Table1 and Figure 1. We printed the main components out by using the SLA 3D printer."),a.default.createElement("table",{className:"table"},a.default.createElement("tbody",null,a.default.createElement("tr",null,a.default.createElement("th",{colSpan:"6",className:"tablehead"},"Peristaltic Pump")),a.default.createElement("tr",null,a.default.createElement("td",{className:"tabledata"},"Dimension"),a.default.createElement("td",{className:"tabledata"},"Tube diameter"),a.default.createElement("td",{className:"tabledata"},"Tube material"),a.default.createElement("td",{className:"tabledata"},"Number of rollers"),a.default.createElement("td",{className:"tabledata"},"Bearing"),a.default.createElement("td",{className:"tabledata"},"Motor")),a.default.createElement("tr",null,a.default.createElement("td",{className:"tabledata"},"31 x 31 x 11 (mm)"),a.default.createElement("td",{className:"tabledata"},a.default.createElement("div",null,"OD: 3mm"),a.default.createElement("div",null,"ID: 2mm")),a.default.createElement("td",{className:"tabledata"},"Silicone rubber"),a.default.createElement("td",{className:"tabledata"},"three"),a.default.createElement("td",{className:"tabledata"},a.default.createElement("div",null,"OD: 9mm"),a.default.createElement("div",null,"ID: 3mm")),a.default.createElement("td",{className:"tabledata"},a.default.createElement("div",null,"28BYJ-48"),a.default.createElement("div",null,"12V"))))),a.default.createElement("div",{className:"undertext"},"Table 1"),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.sampling.pump}),a.default.createElement("div",{className:"undertext"},"Figure 1"))),a.default.createElement("div",{className:"part"},"Because peristaltic pumps deliver fixed amounts of fluid with each pass of a roller over the tube, the size of the tube has a direct effect on the amount of fluid delivered. Well-designed pumps are engineered to work with an optimum tubing size or range of tubing sizes, which take into account both the tube's inside diameter and wall thickness. The inside diameter determines the amount of fluid delivered with each turn of the rotor, while the wall thickness affects the tubing's ability to spring back to its original shape after each compression, which has a great influence on the overall life of the tubing[3]."),a.default.createElement("div",{className:"part"},"We choose the tube with inner diameter 2mm and wall thickness 1mm. Three bearings are used as the rollers in our peristaltic pump. The outer diameter of the bearing is 9mm and the inside diameter is 3mm. The peristatic pump after assembling can be seen in Figure 2. We can also calculated the theoretical discharge to know the theoretical performance of the peristaltic pump by using the equation below[4],"),a.default.createElement("div",{className:"part",style:{display:"flex",alignItems:"center"}},a.default.createElement("div",null,a.default.createElement("div",null,a.default.createElement(s.default.Provider,null,a.default.createElement(s.default.Node,{inline:!0,formula:"flowrate (μl/s) = V \\cdot L \\cdot N \\cdot RPM \\cdot10^3 \\cdot 60"}))),a.default.createElement("div",null,"V= Volume of occluded tubing (mm^3/mm)"),a.default.createElement("div",null,"L= Tubing length that will be occluded by pump rollers (mm)"),a.default.createElement("div",null,"N= Number of rollers on the rotor."),a.default.createElement("div",null,"RPM= pump rpm")),a.default.createElement("div",{style:{marginLeft:"auto",padding:"0 20px"}},a.default.createElement("img",{style:{height:"180px"},src:o.default.image.sampling.full_pump}),a.default.createElement("div",{className:"undertext"},"Figure 2"))),a.default.createElement("div",{className:"part"},a.default.createElement("div",null,"We have done multiple testing and calculated the weight of each droplet by measuring the weight of every ten droplets to get the average value. The average weight of each droplet is 0.035+-0.00072μl, and the rest of the measuring records are showed in Figure 3. This is exactly what we need for sample preparing for the MV-kit in our experiments."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.sampling.list}),a.default.createElement("div",{className:"undertext"},"Figure 3"))),a.default.createElement("div",{className:"part",style:{display:"flex",alignItems:"center"}},a.default.createElement("div",{style:{marginRight:"auto",padding:"0px 30px"}},a.default.createElement("img",{style:{height:"180px"},src:o.default.image.sampling.head}),a.default.createElement("div",{className:"undertext"},"Figure 4")),a.default.createElement("div",null,"Although we can almost get the precise amount of sample each time, it’s still hard to control the later droplets from dropping down cause of the gravity. Hence, we combine the pipette tip (Figure 4) at the end of the silicon tube to reduce the diameter and slow down the flowing rate. In addition, we also design the special lid with slopes in between to cover on the 96-well plate. We can utilize the slope design not only to drop the first few droplets that might be contaminated by the previous sample but also use them as the tunnels to collect the waste sample and remove all of them after the whole examination is done.")),a.default.createElement("div",{className:"illustration"},a.default.createElement("video",{className:"illust-image",autoPlay:!0,muted:!0,loop:!0},a.default.createElement("source",{src:o.default.video.lid,type:"video/mp4"})))),f=a.default.createElement("div",null,a.default.createElement("div",null,"We use two 12V 28BYJ-48 stepper motors to control the XY axis to move our sample to the specific coordinate and connect them to Arduino as well as uln2003 motor drivers to control the speed and time. The step angle of the motor is 5.625 degrees and the reduction ratio is 1/64, therefore, for one revolution it needs 4096 steps. The diameter of the gear mounted on the motor is 36mm, so for each step, it can linearly move forward or backward 1.76mm. The length between two wells on 96-well plate is about 18mm, thus, this motor can absolutely satisfy our need for our low-cost XY plane setting. "),a.default.createElement("div",{className:"illustration"},a.default.createElement("video",{className:"illust-image",autoPlay:!0,muted:!0,loop:!0},a.default.createElement("source",{src:o.default.video.sampling,type:"video/mp4"})))),d=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},"At first, we’re trying to set up the XY axis by using the DC motor to control it in considering of the low-cost budget. However, we found out that since the DC motor is categorized as analog motor, we can only drive them by sending power signals (two wire) to the motor to drive them. This would be tricky for us to do the sophisticated planned moves because we won’t know exactly where they may be located except trying to predict the moves over time within a relative margin of error(3-8%)[5]. Moreover, the friction between the gear rack and the motor make it even hard to control and need to build up more complex moves manually. "),a.default.createElement("div",{className:"part"},"On the other hand, stepper motors are digital motors, we can actually send position signals (four wire) to the motor to drive them. It’s possible for us to know exactly where they are at all times, and would be easier for us to do ‘repeat’ motion passes and have it return to a ‘home and start’ position precisely. For this reason even most sophisticated of a moves can be achieved. "),a.default.createElement("div",{className:"part"},"Due to all the reasons above, instead of making motion decision ‘on the fly’ at the rig, we then choose stepper motor as our driving motor.")),p=a.default.createElement("div",null,a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.sampling.result}))),h=a.default.createElement("div",null,"You can download the STL and anrdion code on ",a.default.createElement("a",{href:"https://github.com/igem-tw-nthu/Peristaltic-pumps-2018"},"Github")),m=a.default.createElement("div",null,a.default.createElement("div",{className:"reference"},"1. https://www.manufacturingchemist.com/news/article_page/Peristaltic_pumps__advantages_and_applications/74693"),a.default.createElement("div",{className:"reference"},"2. Peristaltic pump design: https://www.youtube.com/watch?v=J2KLq7485kM"),a.default.createElement("div",{className:"reference"},"3. https://archive-resources.coleparmer.com/TechInfo/HTMLFiles/PPTube_match.htm"),a.default.createElement("div",{className:"reference"},"4. Suhas R. Dhumal, Design and Development of Rotary Peristaltic Pump, International Journal of Science and Advanced Technology (ISSN 2221-8386)"),a.default.createElement("div",{className:"reference"},"5. https://support.dynamicperception.com/hc/en-us/articles/202986595-What-is-the-difference-between-DC-motors-and-Stepper-motors-")),g={viewbar:{Title:"Sampling",Subtitle:"Auto sampling with peristaltic pump ",image:o.default.image.topfull.sampling,Underimage:"reference: http://keni.candlecomfortzone.com water-samples "},content:{Introduction:"In our device, in order to let the following detected process work out properly, we first set up the automated sampling mechanism. 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However, we found that it is not bright enough, so we chose another kind of light."),a.default.createElement("div",{className:"part"},"After this, we choose the white light bar and connect it with Arduino. Since the voltage afforded by Arduino is 12V, it’s too bright to observe, so we apply the voltage-gain-module to get the appropriate voltage. Then we find out the white light may sometimes be difficult for us to tell apart whether the liquid changes color or not. After discussion, we change the white light to yellow light and the problem is solved. ")),c=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},'Our Vibrio detection device needs to be able to react in a stable 30 ° C environment. It is hoped that a method can be designed to heat up when the ambient temperature is high and cool down when the temperature is low. The cooling technologies used today are mainly the following: "high pressure gas expansion and cooling", "liquid evaporation and refrigeration", "gas vortex cooling", "semiconductor refrigeration" and "chemical cooling", and we choose "semiconductor cooling", which is a relatively easy to complete and within the capability range.'),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"First period"),a.default.createElement("div",{className:"innerpart"},"Common methods for cooling semiconductors include water cooling and fan cooling. Among them, due to our large architecture, water cooling is effective but too cumbersome and heavy. Limited by the space configuration, we use fan cooling. (Picture 4) By keep bringing the heat away, the temperature gradient of the chip become larger, and finally it will stop at a balanced point. We also add a fan on the cold side since we want the cold air to be uniformed inside the box. (Picture 5) Temperature-controlled circuits are more cumbersome and too complicated circuits are not suitable for use in our system, so we try to use temperature control module first. (Picture 6)"),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:l.default.image.environment.first}),a.default.createElement("div",{className:"undertext"})),a.default.createElement("div",{className:"innerpart"},"We found that as we put the cooling device in a cardboard box for testing, the result is not good since the space is not sealed and made the chip hard to cool down the environment."),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",{className:"innerpart"},"Needed materials:"),a.default.createElement("div",null,"two 5V-2wire fans, two small heat sinks, some thermal grease, one thermoelectric cooling chip provided by a national competition in Taiwan, temperature control module."))),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Second period"),a.default.createElement("div",{className:"innerpart"},"To enlarge the temperature difference, we add a more powerful fan on the hot side to make sure the rate of heat dissipation is enough for the rate of chip getting cold, which is a 12V-2wire one. As a result, the inbox environment could drop 3℃, but still didn’t reach our expectation."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:l.default.image.environment.second}),a.default.createElement("div",{className:"undertext"},a.default.createElement("div",null,"Figure 1: The air temperature inside the box drops from 26.6℃ to about 18℃ in 7 minutes, and maintains at 18℃ for 30 minutes. (cold device) The air temperature inside the box rises from 27.2℃ to 32.8℃ in about 6 minutes, and maintains at 32.8℃ for 15 minutes. (hot device)"),a.default.createElement("div",null,"Figure 2: When cold devices turn off, the air temperature inside the box rises from 27.2℃ to 32.8℃ in about 6 minutes, and maintains at 32.8℃ for 15 minutes. (cold device) When the hot device turn off, the air temperature inside the box drops from about 32.8℃ to 27.6℃ in 6 minutes, the same as the time to heat up the air. (hot device)"))),a.default.createElement("div",{className:"innerpart"},'During our experiment, we found that the temperature control module couldn’t provide a two-state system, which is hard to be designed in our project. Therefore, we try another way by using Arduino and relay module to control the temperature directly. Relay, also known as electric cymbal, is an electronic control device that has a control system (also known as an input circuit) and a controlled system (also known as an output circuit). It is usually used in automatic control circuits. It is actually used as an "automatic switch" that uses a small current to control large currents. Therefore, it plays the role of automatic adjustment, safety protection and conversion circuit in the circuit. Besides, we chose an IC called LM35 as our temperature sensor by transforming resistance into temperature. The wiring diagram is as below. (Picture 8)'),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",{className:"innerpart"},"Needed materials: "),"Cold device is combined by 12V-2wire fan, heat sink, thermal grease, thermoelectric cooling chip, thermal grease, heat sink, 5V-2wire fan (from up to down); Temperature controlling: Arduino for control, LM35 for sensing, relay module for switch.")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Final period"),a.default.createElement("div",{className:"innerpart"},"To reduce the unknown impact on the experiment, we use two cold device and one hot device to control the temperature. Hot device is similar to cold one, the only different is its two fans are all 5V-2wire, which is the device we used in the first period."),"  ",a.default.createElement("div",{className:"innerpart"},"This time we use two cold devices to find for the lowest temperature. Figure 1 shows the result we test cold or hot devices, and Figure 2 shows the changing after we turn off the devices."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:l.default.image.environment.final}),a.default.createElement("div",{className:"undertext"})),a.default.createElement("div",{className:"inner-part"},"After discussion, we decide to give styrofoam a try, we add it inside the box and do the rest of the experiment, includes add it or not add it. We finally decide to use two cold devices and one hot device without adding any material."),a.default.createElement("table",null),a.default.createElement("div",{className:"innerpart"},"We use Arduino connects with relay and temperature sensor to control the temperature. When the sensor detects the temperature lower than the setting, hot device is activated, it would heat up the air inside the device until it reaches the target temperature. Then, the hot device continues working that makes the temperature higher than the ideal one, this time, the cold devices turn on and cool down the environment. At the end, the temperature would be controlled in the range we have written in Arduino."),a.default.createElement("div",{className:"innerpart"},"In the beginning, we set the temperature to 27℃, which is a good condition for germs to grow. However, we neglect that there would be some the delay of temperature change between the air and the sensor, and the temperature around the sensor became higher than the environment, which affected the detecting correction. After we tidied up the wires, the problems disappeared and came into perfect."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:l.default.image.environment.end}),a.default.createElement("div",{className:"undertext"},"Figure 6: Temperature control simulation. The hot device turns on at the beginning, and the cold devices turn on when the temperature reaches 28.3℃, in about 6 minutes.")))),f=a.default.createElement("div",null,a.default.createElement("div",{className:"reference"},"1. Thermal Conductivity and Thermoelectric Power of Semiconductors. I. Terasaki, in Reference Module in Materials Science and Materials Engineering, 2016"),a.default.createElement("div",{className:"reference"},"2. Physics and Fundamental Theory. I. Terasaki, in Comprehensive Semiconductor Science and Technology, 2011"),a.default.createElement("div",{className:"reference"},"3. Thermoelectric cooling. JoséMeseguer, IsabelPérez-Grande, AngelSanz-Andrés, in Spacecraft Thermal Control, 2012"),a.default.createElement("div",{className:"reference"},"4. https://en.wikipedia.org/wiki/Thermoelectric_effect#/media/File:Thermoelectric_Cooler_Diagram.svg")),d={viewbar:{Title:"Environment",Subtitle:"Led Light, and peltier cooler",image:l.default.image.topfull.environment},content:{Introduction:"In order to make sure the observation results are stable and reliable, it’s necessary to control the experiment environment. Besides putting all the instruments inside our acrylic-made box (size: 20*30*35, unit: cm3), temperature and brightness are also important. In this case, we separate the “Environment” part into two sub-titles, temperature, and light. We will introduce more details in the following statements.",parts:[{Title:"Light",Description:u},{Title:"Temperature",Description:c},{Title:"Reference",Description:f}]}},p=function(e){function t(e){return function(e,t){if(!(e instanceof t))throw new TypeError("Cannot call a class as a function")}(this,t),function(e,t){if(!e)throw new ReferenceError("this hasn't been initialised - super() hasn't been called");return!t||"object"!=typeof t&&"function"!=typeof t?e:t}(this,(t.__proto__||Object.getPrototypeOf(t)).call(this,e))}return function(e,t){if("function"!=typeof t&&null!==t)throw new TypeError("Super expression must either be null or a function, not "+typeof t);e.prototype=Object.create(t&&t.prototype,{constructor:{value:e,enumerable:!1,writable:!0,configurable:!0}}),t&&(Object.setPrototypeOf?Object.setPrototypeOf(e,t):e.__proto__=t)}(t,i.Component),r(t,[{key:"render",value:function(){return a.default.createElement(o.default,d)}}]),t}();t.default=p},function(e,t,n){"use strict";Object.defineProperty(t,"__esModule",{value:!0});var r=function(){function e(e,t){for(var n=0;n<t.length;n++){var r=t[n];r.enumerable=r.enumerable||!1,r.configurable=!0,"value"in r&&(r.writable=!0),Object.defineProperty(e,r.key,r)}}return function(t,n,r){return n&&e(t.prototype,n),r&&e(t,r),t}}(),i=n(0),a=s(i),o=s(n(4)),l=s(n(8));function s(e){return e&&e.__esModule?e:{default:e}}var u=a.default.createElement("div",null,a.default.createElement("div",null,"In order to achieve real time data analysis , when ACE detects the color-changing sample, it will send a message to the user. Between ACE and user’s mobile device, it needs a transfer protocol to communicate with each other. Here, we choose MQTT (Message Queuing Telemetry Transport) to upload the vibrio concentration onto the cloud. It is a lightweight messaging protocol for small sensors and mobile devices, optimized for high-latency or unreliable networks.  Therefore, this kind of method is useful in remote area where network bandwidth is limited, especially for the fish farm in Taiwan. We’ve designed an app for farmers to obtain information in time. In our app, we separate into three parts: Pond’s condition, Suggestions, and Multiple Pond map. "),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.software.app}))),c=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},"We’ve divided different bacteria’s concentration into three levels, Safe, Warning, and Danger. The standard of this division is refer to the advice of the Fisheries Research Center of Republic of China. After visiting the Marine Life museum Research center, the researcher also tell us that usually the concentration for vibrio to be pathogenic is >104CFU/ml."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.software.line})),a.default.createElement("div",{className:"part"},"Furthermore, in order to let the farmers keep track of how the vibrio in their farm has been growing, we’ve made a history record chart, of which the Y coordinate is concentration, and the X coordinate is time. The concentration points will be drawn every hour.  The chart can be drag, zoom in and out for a closer look. By knowing the condition of their pond beforehand and even predict how the amount of the vibrios might grow, the farmers can do precaution in order to prevent vibrios from harming their fish or shrimp.")),f=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},"When the concentration of vibrios is under the safety standard, the fisheries research center recommend the farmers to change the water environment, or adjust the amount of feed. Only when the condition is in danger that they can add the medicine.  "),a.default.createElement("div",{className:"part"},"In addition, we will send notification bar through android phones to farmers if the amount of vibrios exceed standard."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.software.warn}))),d=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},"The reason why we want to combine google map into our APP is that one farmer usually own more than one pond.  Then if they want to keep track of different pond, they need to number their farm. Most of the time, it’s difficult for farmers to memorize which pond does this number really referring to, which indicate that the managing system is not so user friendly. "),a.default.createElement("div",{className:"part"},"In our map, farmers can locate its position and to help its pond create a name.  Next time when they want to inspect the condition of each pond, they can just press on the name label on the map. In order to make the system more intuitive, we add in the function of turning the normal map into 3D map and satellite(RealView) one, which is more close to the real view on the ground."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.software.map}))),p={viewbar:{Title:"Software",Subtitle:"An app for real-time analysis of pond’s condition",image:o.default.image.topfull.software},content:{Introduction:u,parts:[{Title:"Pond’s condition",Description:c},{Title:"Suggestions",Description:f},{Title:"Multiple pond management map",Description:d}]}},h=function(e){function t(){return function(e,t){if(!(e instanceof t))throw new TypeError("Cannot call a class as a function")}(this,t),function(e,t){if(!e)throw new ReferenceError("this hasn't been initialised - super() hasn't been called");return!t||"object"!=typeof t&&"function"!=typeof t?e:t}(this,(t.__proto__||Object.getPrototypeOf(t)).apply(this,arguments))}return function(e,t){if("function"!=typeof t&&null!==t)throw new TypeError("Super expression must either be null or a function, not "+typeof t);e.prototype=Object.create(t&&t.prototype,{constructor:{value:e,enumerable:!1,writable:!0,configurable:!0}}),t&&(Object.setPrototypeOf?Object.setPrototypeOf(e,t):e.__proto__=t)}(t,i.Component),r(t,[{key:"render",value:function(){return a.default.createElement(l.default,p)}}]),t}();t.default=h},function(e,t,n){"use strict";Object.defineProperty(t,"__esModule",{value:!0});var r=function(){function e(e,t){for(var n=0;n<t.length;n++){var r=t[n];r.enumerable=r.enumerable||!1,r.configurable=!0,"value"in r&&(r.writable=!0),Object.defineProperty(e,r.key,r)}}return function(t,n,r){return n&&e(t.prototype,n),r&&e(t,r),t}}(),i=n(0),a=s(i),o=s(n(8)),l=s(n(4));function s(e){return e&&e.__esModule?e:{default:e}}var u=a.default.createElement("div",null,a.default.createElement("div",null,"The importance of damage caused by Vibrio would not be elucidated here. You can see the detailed information in",a.default.createElement("a",{href:"https://2018.igem.org/Description"}," Description"),"."),a.default.createElement("div",{className:"part"},a.default.createElement("div",null,"Nowadays, there are three possible scenarios for aquatic agriculture to face pathogenic Vibrio:"),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",null,"1. The infection of Vibrio is usually happening all of a sudden. So when fisherman aware of the infection, it often causes significant damage."),a.default.createElement("div",null,"2. The conventional Vibrio testing method is time-consuming and manpower-consuming."),a.default.createElement("div",null,"3. The current water quality testing equipment is mainly focusing on the physical and chemical index, there are no devices for testing bacteria"))),a.default.createElement("div",{className:"part"},a.default.createElement("div",null,"However, our automatic Vibrio-detecting system is able to solve these problems."),a.default.createElement("div",{style:{display:"flex",alignItems:"flex-end"}},a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:l.default.image.application.tcbs}),a.default.createElement("div",{className:"undertext"},"TCBS medium")),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:l.default.image.application.plate}),a.default.createElement("div",{className:"undertext"},"MV-kit"))))),c=a.default.createElement("div",null,a.default.createElement("div",null,"MV-kit was developed by Fisheries Research Institute in Taiwan. It’s a kind of selective medium for Vibrio and it can detect 17 different aquatic pathogenic Vibrio. Through observing the time of color-changing, we can know the approximate Vibrio concentration in the sample. After our modification, within 24 hours, the minimal detectable Vibrio concentration is 100 CFU/ mL. Take Vibrio harveyi for example:"),a.default.createElement("div",{style:{display:"flex",alignItems:"flex-end"}},a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:l.default.image.application.concentration}),a.default.createElement("div",{className:"undertext"},"CFU vs. O.D.")),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:l.default.image.application.time}),a.default.createElement("div",{className:"undertext"},"CFU vs. Time of color-changing"))),a.default.createElement("div",null,"We build a relationship of O.D. value and the amount of bacteria via spreading method. Then, using O.D. value as a reference point, we build another relationship between the color-changing time of MV-kit and the amount of bacteria. Accordingly, we can get a significant correlation of bacteria concentration and the color-changing time of MV-kit")),f=a.default.createElement("div",null,a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:l.default.image.application.flow})),a.default.createElement("div",{className:"part"},"We provide a complete service for the prevention of Vibrio-infection. Our ACE collects the water sample from the fish pond and drips it into the plate automatically. Simultaneously, the device provides a closed environment with constant temperature for the reaction. Also, the device monitors the color-changing time at all times. As long as the color changes and achieves the condition we have set up, the data will be recorded and be uploaded to the server. After analyzing, the final result will be transferred to the clients’ cellphone applications. If the concentration of Vibrio overtakes the minimal hazardous value which is 1000 CFU/ mL, the application will warn the fisherman."),a.default.createElement("div",{className:"part"},"Through our system, our clients can take control of the situation of their fish ponds and use our engineered E. coli or other plans to solve the problems.")),d=a.default.createElement("div",null,a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:l.default.image.application.stp})),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"1. Segmentation"),a.default.createElement("div",null,a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",{className:"part-title"},"Merchandises in current market have to face some difficulties:"),a.default.createElement("div",null,"The water quality detecting systems in the current market are merely focusing on the particular physical and chemical indexes in the waters, for example, pH value, redox potential, dissolved oxygen, etc. They determine the water quality by those indexes, however, the change of situation in the water is quite fast and complex. We cannot receive the correct and real-time information of Vibrio in the water. When we aware of the abnormality, the pathogenic Vibrio usually causes the death of fish and shrimps significantly.")),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",{className:"part-title"},"Long-term detection with high specificity:"),a.default.createElement("div",null,"The MV-kit developed by Fisheries Research Institute is able to detect the Vibrio in the water sample quantitatively and with high specificity. Our system can automatically monitor the change of Vibrio concentration over a long period of time. Our clients are able to notice the condition in the early stage of Vibrio growth and adopt the solutions.")),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",{className:"part-title"},"A real-time and automatic system:"),a.default.createElement("div",null,"Our mechanism design allows the device to collect the sample automatically and continuously. Furthermore, we also program an application which analyzes the Vibrio concentration automatically. Last but not least, we achieve the effect of real-time monitoring and report through IoT technology.")),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",{className:"part-title"},"Fast, simple, and low-cost:"),a.default.createElement("div",null,"The mechanism of our reaction is the simple color-changing process. Compared to the conventional Vibrio culture methods, our clients haven’t to equipped with the professional skills of bacteria culture, also, the process of testing becomes safer and faster. In comparison with the water quality testing system in the market, our system is low-cost and is able to express the Vibrio concentration directly and accurately."))),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:l.default.image.application.vs}))),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"2. Targeting"),a.default.createElement("div",null,a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",null,"Aquatic agriculture industry:"),a.default.createElement("div",null,"Aquatic agriculture is an important industry and it stimulates the economic growth in Taiwan every year. However, the Vibrio-related infection leads to inestimable economic damage locally and globally. Our system aims to provide real-time, long-term, and low-cost monitoring toward Vibrio. Through the automatic system, our clients can reduce the manpower-cost and ensure their fish ponds are safe at any time. In addition, our mechanism design is quite simple so that the maintenance cost is also quite low. The only thing fisherman has to do is change the consumable materials regularly.")),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",{className:"part-title"},"Suppliers of the water quality testing system:"),a.default.createElement("div",null,"Currently, there are several water quality testing system vendors in Taiwan, but there is no any company provides the automatic Vibrio-detecting system. Our device is easy to be integrated with other related systems to reduce the vendors’ research and development cost. In this way, we believe that we can establish a better platform and to create a win-win situation.")))),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"3. Positioning"),a.default.createElement("div",null,a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",{className:"part-title"},"Intelligent fishery:"),a.default.createElement("div",null,a.default.createElement("div",null,"With the advance of science and technology, the IoT popularizes gradually in current society. Besides, the traditional agriculture industries transform into the so-called “agriculture 4.0”. "),a.default.createElement("div",null,"Let’s take an in-depth look at aquatic agriculture in Taiwan. Our country is famous for its aquatic agriculture industry, however, the conventional way is quite time-consuming. Additionally, following with the global warming, the aquatic agriculture faces a huge challenge. In the train of those facts, the industrial population structure becomes imbalance and thus, exerting a crucial impact on the future of industrial development ultimately."),a.default.createElement("div",null,"In order to turn the tables, the concept and development of intelligent fishery are emerging in recent years. It leads our traditional fishery industry into a new era, raising the operational efficacy, the income, and reducing the cost. Based on the concept, our system is expected to solve the Vibrio-monitoring problems and to establish a database to find an optimized solution."))),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",{className:"part-title"},"Future resolution:"),a.default.createElement("div",null,"Hopefully, we can create a complete detecting platform for different pathogenic bacteria in the future. With low-cost and high-efficiency, our platform is able to minimize the economic damage result from the bacterial infection."))))),p={viewbar:{Title:"Application",Subtitle:"Automatic Vibrio-detecting System",image:l.default.image.topfull.application},content:{Introduction:u,parts:[{Title:"MV-kit",Description:c},{Title:"Automatic Vibrio-detecting System",Description:f},{Title:"Commercial Analysis",Description:d}]}},h=function(e){function t(){return function(e,t){if(!(e instanceof t))throw new TypeError("Cannot call a class as a function")}(this,t),function(e,t){if(!e)throw new ReferenceError("this hasn't been initialised - super() hasn't been called");return!t||"object"!=typeof t&&"function"!=typeof t?e:t}(this,(t.__proto__||Object.getPrototypeOf(t)).apply(this,arguments))}return function(e,t){if("function"!=typeof t&&null!==t)throw new TypeError("Super expression must either be null or a function, not "+typeof t);e.prototype=Object.create(t&&t.prototype,{constructor:{value:e,enumerable:!1,writable:!0,configurable:!0}}),t&&(Object.setPrototypeOf?Object.setPrototypeOf(e,t):e.__proto__=t)}(t,i.Component),r(t,[{key:"render",value:function(){return a.default.createElement(o.default,p)}}]),t}();t.default=h},function(e,t,n){"use strict";Object.defineProperty(t,"__esModule",{value:!0});var r=function(){function e(e,t){for(var n=0;n<t.length;n++){var r=t[n];r.enumerable=r.enumerable||!1,r.configurable=!0,"value"in r&&(r.writable=!0),Object.defineProperty(e,r.key,r)}}return function(t,n,r){return n&&e(t.prototype,n),r&&e(t,r),t}}(),i=n(0),a=s(i),o=s(n(8)),l=s(n(4));function s(e){return e&&e.__esModule?e:{default:e}}var u=a.default.createElement("div",null,a.default.createElement("div",null,"In order to control Vibrio cell-population density and maintain a bio-friendly environment in a sustainable way, we introduce the concept of biocontrol, which is known as the prey-predator model, to our system. We create a system that can automatically detect, compare, and control the cell-population density. The system is composed of three parts, which are a detector, comparator, and controller respectively (Fig 1). These three parts are connected together in the same gene circuit and function sequentially so that the concentration of Vibrio in the surroundings can be kept in a specific range. Then, a succession model can be achieved."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:l.default.image.wetlab_overview.system}),a.default.createElement("div",{className:"undertext"},"Fig 1. Component of cell population controlling system."))),c=a.default.createElement("div",null,"Our gene circuit design can be divided into three parts described down below."),f=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},"The Detector module is responsible for detecting the concentration of Vibrio species in the surroundings. We found that most of pathogenic Vibrio use quorum sensing to communicate with other individuals. Lux is one of the Vibrio quorum sensing systems. With Lux system, the Vibrio has the ability to secrete and detect specific chemicals, acyl homoserine lactones (AHLs, Fig2). Signal transduction relies on AHLs, and bacteria respond to it with different gene expression."),a.default.createElement("div",{className:"part"},"We use Lux gene originally isolated from Vibrio fischeri as our AHL sensor. The gene system is composed of LuxI, LuxR, and pLux. LuxI is a synthase that converts S-Adenosyl methionine (SAM) into one of specific AHLs, 3OC6HSL (Fig2). LuxR is an activating protein, and it will stimulate the transcription from Lux promoter (pLux) when 3OC6HSL binds to it.[1] We modify LuxI with RFP reporter to the E. coli. playing the role of Vibrio. We also modify LuxR and pLux into our genetically engineered E. coli., the cell-population controlling system, so that AHL level in the environment can be detected and the Vibrio population can be known (Fig3).        "),a.default.createElement("div",{className:"illustration"},a.default.createElement("div",{className:"illust-image-wrapper"},a.default.createElement("img",{className:"illust-image",src:l.default.image.wetlab_overview.ahl})),a.default.createElement("div",{className:"undertext"},"Fig 2. Chemical structure of 3-oxo-C6-HSL.")),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:l.default.image.wetlab_overview.detection}),a.default.createElement("div",{className:"undertext"},"Fig 3. An AHL detection scheme showing that AHL-LuxR complex activates the gene downstream from pLux."))),d=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},"The comparator module is responsible for comparing the cell population density of Vibrio with our engineered E. coli. to see if it exceeds the pathogenic level. It receives information getting from the detector module. If the concentration of Vibrio is higher than engineered E. coli., which means LuxR-AHL complex is present, the anti-Vibrio gene will be transcribed. On the other hand, if the concentration of engineered E. coli. is higher than Vibrio, which means LuxR-AHL complex is absent, the suicide gene will be transcribed to kill E. coli. itself. "),a.default.createElement("div",{className:"part"},"Our initial effort is to design a gene circuit contain LacR to accomplish our goal. However, it turns out that there are some problems and drawbacks. While AHL presences, LacR should be produced and bound to pLac promoter to inhibit suicide gene being expressed. Yet, LacR can’t 100 % bind to pLac and this happens in most operons, so-called leakage problem. Suicide gene may be expressed, and E. coli. will suicide even at high AHL concentration. This would cause the system to be collapsed, and the result became out of our expectations (Fig 4)."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:l.default.image.wetlab_overview.lacl}),a.default.createElement("div",{className:"undertext"},"Fig 4. Our initial design using Lac operon to regulate genes expression.")),a.default.createElement("div",{className:"part"},"We come up with a better solution for our kill switch by using riboswitch and Small Transcriptional-Activating RNA (STAR) to regulate our gene. STAR is a small RNA with its sequence completely complementary to riboswitch. Riboswitch is an intrinsic transcription terminator, a DNA segment with special secondary structure. It forms a hairpin structure and inhibits transcriptions of the genes downstream.[2] When STAR binds to the riboswitch, it will be turned on (Fig 5, 6). Current studies show that riboswitch can regulate protein better than operon because of the higher affinity between STAR and riboswitch. In addition, STAR is smaller than regular proteins and it degrades faster than regular protein, therefore can respond to environmental change more rapidly than operon."),a.default.createElement("div",{className:"illustration"},a.default.createElement("div",{className:"illust-image-wrapper"},a.default.createElement("img",{className:"illust-image",src:l.default.image.wetlab_overview.riboswitch})),a.default.createElement("div",{className:"undertext"},"Fig 5. The secondary structure of riboswitch and STAR")),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:l.default.image.wetlab_overview.star}),a.default.createElement("div",{className:"undertext"},"Fig 6. STAR turns on the target and activates the downstream transcriptional process")),a.default.createElement("div",{className:"part"},"We also use a small RNA segment called anti-STAR in our design. Its sequence is totally complementary to STAR, but has higher affinity than riboswitch does. When anti-SATR exists, STAR will bind to it prior to riboswitch. Consequently, when Lux-AHL triggers the transcription of anti-STAR, it binds to STAR and the riboswitch is turned off (Fig 7). When Lux-AHL is absent and anti-STAR isn’t generated, STAR will turn on the riboswitch."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:l.default.image.wetlab_overview.anti}),a.default.createElement("div",{className:"undertext"},"Fig 7. Anti-STAR makes STAR lose it functionality. The gene downstream is still inhibited by the target."))),p=a.default.createElement("div",null,a.default.createElement("div",null,"We use two growth inhibitors to control populations of Vibrio and engineered E. coli. One is for inhibiting Vibrio, and another is for inhibiting engineered E. coli. At first, we would like to use anti-Vibrio substances in our design. However, our lab belongs to Bio-safety Level 1 (BSL1) and we can’t use real Vibrio in our experiments to test the toxicity of anti-Vibrio substance due to bio-safety concern. Therefore, we use colicin M to kill E. coli. (fake Vibrio) in order to prove our bio-control model. On the other hand, we use toxin small RNA (tsRNA) and endolysin as suicide substances to kill engineered E. coli. itself."),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"1. Colicin M"),a.default.createElement("div",null,"Colicin M blocks the biosynthesis of peptidoglycan (murein) and O-antigen. Colicin M contains activity (cma) and immune gene (cmi). The product of cmi is an immune protein, preventing colicin M to inhibit cell wall synthesis by binding to and thus neutralizing it. In addition, most colicins are released by autolysis, but colicin M lacks a lysis gene.[3-4] It has less tendency to cause the death of producer cells, so it’s a suitable protein for the purpose of killing other bacteria without harming producer cells.")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"2. Toxic small RNA (tsRNA)"),a.default.createElement("div",null,"Toxic small RNAs are small non-coding RNA. We use the tsRNA isolated from Burkholderia cenocepacia. The Current study showed that it is toxic to the E. coli. host and reproducibly inhibit E. coli growth. TsRNA harbored a sequence that was complementary to the consensus ribosomal binding site (RBS). It is hypothesized that it might target and misregulate essential genes within the E. coli cell. TsRNA cannot penetrate the bacterial cell from the outside, so it is unable to affect bacteria except E. coli. host cells.[5] Additionally, producing tsRNA doesn’t require a translational process, thus the response time is more rapid. As a result, tsRNA become an ideal suicide substance for our design.")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"3. Endolysin"),a.default.createElement("div",null,"Endolysin is a peptidoglycan hydrolase that is secreted by double-stranded DNA lambda phage to comprise the bacterial cell wall. It causes cell lysis by disrupting murein structure.")),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:l.default.image.wetlab_overview.mechanism}),a.default.createElement("div",{className:"undertext"},"Fig 8. The overall gene circuit design"))),h=a.default.createElement("div",null,a.default.createElement("div",{className:"innerpart"},"1. Visick, K. L.; Foster, J.; Doino, J.; McFall-Ngai, M.; Ruby, E. G., Vibrio fischeri lux Genes Play an Important Role in Colonization and Development of the Host Light Organ. Journal of Bacteriology 2000, 182 (16), 4578-4586."),a.default.createElement("div",{className:"innerpart"},"2. Chappell, J.; Takahashi, M. K.; Lucks, J. B., Creating small transcription activating RNAs. Nature chemical biology 2015, 11 (3), 214-20."),a.default.createElement("div",{className:"innerpart"},"3. Harkness, R. E.; Olschlager, T., The biology of colicin M. FEMS microbiology reviews 1991, 8 (1), 27-41."),a.default.createElement("div",{className:"innerpart"},"4. Zeth, K.; Römer, C.; Patzer, S. I.; Braun, V., Crystal Structure of Colicin M, a Novel Phosphatase Specifically Imported by Escherichia coli>. The Journal of Biological Chemistry 2008, 283 (37), 25324-25331."),a.default.createElement("div",{className:"innerpart"},"5. Kimelman, A.; Levy, A.; Sberro, H.; Kidron, S.; Leavitt, A.; Amitai, G.; Yoder-Himes, D. R.; Wurtzel, O.; Zhu, Y.; Rubin, E. M.; Sorek, R., A vast collection of microbial genes that are toxic to bacteria. Genome research 2012, 22 (4), 802-9.")),m={viewbar:{Title:"Wetlab",Subtitle:"Detector, comparator, and controller module",image:l.default.image.topfull.wetlab_overview},content:{Introduction:u,parts:[{Title:"Cell Population Controlling System",Description:c},{Title:"Detector",Description:f},{Title:"Comparator",Description:d},{Title:"Controller",Description:p},{Title:"Reference",Description:h}]}},g=function(e){function t(){return function(e,t){if(!(e instanceof t))throw new TypeError("Cannot call a class as a function")}(this,t),function(e,t){if(!e)throw new ReferenceError("this hasn't been initialised - super() hasn't been called");return!t||"object"!=typeof t&&"function"!=typeof t?e:t}(this,(t.__proto__||Object.getPrototypeOf(t)).apply(this,arguments))}return function(e,t){if("function"!=typeof t&&null!==t)throw new TypeError("Super expression must either be null or a function, not "+typeof t);e.prototype=Object.create(t&&t.prototype,{constructor:{value:e,enumerable:!1,writable:!0,configurable:!0}}),t&&(Object.setPrototypeOf?Object.setPrototypeOf(e,t):e.__proto__=t)}(t,i.Component),r(t,[{key:"render",value:function(){return a.default.createElement(o.default,m)}}]),t}();t.default=g},function(e,t,n){"use strict";Object.defineProperty(t,"__esModule",{value:!0});var r=function(){function e(e,t){for(var n=0;n<t.length;n++){var r=t[n];r.enumerable=r.enumerable||!1,r.configurable=!0,"value"in r&&(r.writable=!0),Object.defineProperty(e,r.key,r)}}return function(t,n,r){return n&&e(t.prototype,n),r&&e(t,r),t}}(),i=n(0),a=u(i),o=u(n(4)),l=u(n(8)),s=u(n(136));function u(e){return e&&e.__esModule?e:{default:e}}var c=a.default.createElement("div",null,a.default.createElement("div",{className:"illustration"},a.default.createElement("div",{style:{margin:"auto",width:"500px"}},a.default.createElement("img",{className:"illust-image",src:o.default.image.model.interaction}))),a.default.createElement("div",{className:"part"},"We genetically modify the E.coli, giving them the ability to detect Vibrio concentration, and secrete some compound to supress Vibrio and itself. "),a.default.createElement("div",{className:"part"},"If the AHL concentration exceed a specific threshold, engineered E.coli will secrete antibacterial peptide Colisin M to kill Vibrio. Otherwise, the Vibrio concentration below the E.coli's detection threshold, engineered Ecoli will secrete tsRNA to commit suicide.")),f=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},a.default.createElement("div",null,"We revise this model from Lotka-Volterra model with logistic population groth, and the decay rate of compound."),a.default.createElement("div",{style:{width:"fit-content",margin:"auto"}},a.default.createElement(s.default.Provider,null,a.default.createElement("div",{style:{marginTop:"20px"}},a.default.createElement(s.default.Node,{inline:!0,formula:"{{dV} \\over{dt}} = r_v V - {{r_v} \\over {V_{max}}} V^2 -a VN"})),a.default.createElement("div",{style:{marginTop:"20px"}},a.default.createElement(s.default.Node,{inline:!0,formula:"{{dA} \\over {dt}} = k_AV - \\lambda_A A"})),a.default.createElement("div",{style:{marginTop:"20px"}},a.default.createElement(s.default.Node,{inline:!0,formula:"{{dE} \\over {dt}} = bEA - cES - \\lambda_E E"})),a.default.createElement("div",{style:{marginTop:"20px"}},a.default.createElement(s.default.Node,{inline:!0,formula:"{{dC} \\over {dt}} = k_CE - \\lambda_C C "})),a.default.createElement("div",{style:{marginTop:"20px"}},a.default.createElement(s.default.Node,{inline:!0,formula:"{{dS} \\over {dt}} = k_SE - \\lambda_S S"}))))),a.default.createElement("div",{className:"part"},"With the contraint of AHL threshold",a.default.createElement("div",{style:{width:"fit-content",margin:"auto"}},a.default.createElement(s.default.Provider,null,a.default.createElement("div",{style:{marginTop:"20px"}},a.default.createElement(s.default.Node,{inline:!0,formula:"A \\lt A_t, \\ k_C=0"})),a.default.createElement("div",{style:{marginTop:"20px"}},a.default.createElement(s.default.Node,{inline:!0,formula:"A \\geq A_t, \\ k_S=0"})))))),d=a.default.createElement("div",null,"With changing specific parameter, such as binding strength of antibacterial Colisin with Vibrio, binding strength of Ahl with Ecoli, and the detection threshold of AHL for engineered E.coli. We are able to simulate the population of two bacteria in oscillating, and a parameter sensitive constant mode.",a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.model.result}),a.default.createElement("div",{className:"undertext"},"Two figure are just a slight change of the Binding strength of Ahl with Ecoli"))),p=a.default.createElement("div",null,"Code detail On ",a.default.createElement("a",{href:"https://github.com/igem-tw-nthu/model-2018"},"Github")),h={viewbar:{Title:"Model",Subtitle:"Build a simple bio-oscillator",image:o.default.image.topfull.model},content:{Introduction:"This year 2018 iGEM NTHU_Taiwan aims to solve the Vibrio infection in aquaculture with bio-competiotion. We are inspired by the peroidic pattern of Lotka-Volterra model, and try to control the Vibrio population with engineered E.coli.",parts:[{Title:"Interaction",Description:c},{Title:"Equations",Description:f},{Title:"Discussion",Description:d},{Title:"Soucre Code",Description:p}]}},m=function(e){function t(){return function(e,t){if(!(e instanceof t))throw new TypeError("Cannot call a class as a function")}(this,t),function(e,t){if(!e)throw new ReferenceError("this hasn't been initialised - super() hasn't been called");return!t||"object"!=typeof t&&"function"!=typeof t?e:t}(this,(t.__proto__||Object.getPrototypeOf(t)).apply(this,arguments))}return function(e,t){if("function"!=typeof t&&null!==t)throw new TypeError("Super expression must either be null or a function, not "+typeof t);e.prototype=Object.create(t&&t.prototype,{constructor:{value:e,enumerable:!1,writable:!0,configurable:!0}}),t&&(Object.setPrototypeOf?Object.setPrototypeOf(e,t):e.__proto__=t)}(t,i.Component),r(t,[{key:"render",value:function(){return a.default.createElement(l.default,h)}}]),t}();t.default=m},function(e,t,n){"use strict";Object.defineProperty(t,"__esModule",{value:!0}),t.default=void 0;var r=function(){function e(e,t){for(var n=0;n<t.length;n++){var r=t[n];r.enumerable=r.enumerable||!1,r.configurable=!0,"value"in r&&(r.writable=!0),Object.defineProperty(e,r.key,r)}}return function(t,n,r){return n&&e(t.prototype,n),r&&e(t,r),t}}(),i=n(0),a=s(i),o=s(n(4)),l=s(n(8));function s(e){return e&&e.__esModule?e:{default:e}}var u=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Transformation"),a.default.createElement("div",null,"During this summer, we had done lots of digestion-ligation and transformation experiments! We enjoyed and had fun exploring the mystery of synthetic biology!"),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.experiment.transform}),a.default.createElement("div",{className:"undertext"},"Fig. 1: Successful transformation of part BBa_K2774008 (Left) and BBa_K2774005 (Right)")),a.default.createElement("div",null,"Utilizing iGEM distribution kit and parts from the registry, we assembled various bio-bricks to construct our genes. We constructed our part, BBa_K2774008, by using pLac with RBS (BBa_J04500), endolysin (BBa_K1378032) with GFP (E0240). The left LB plate was the transformation of BBa_K2774008. The transformation was success and therefore some of the colony emits GFP fluorescence! And the right LB plate was the transformation of part BBa_K2774005, the RFP shows the right colony with successful ligation plasmid. In addition, thanks for the established technology of IDT (Integrated DNA Technologies), providing us the gene fragments. Finally, we successfully constructed tsRNA (BBa_K2774001).")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Electrophoretogram"),a.default.createElement("div",null,"Here are the parts we assemble successfully. We use VF2 and VR primer to check our gene’s length, all of them are on pSB1C3."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.experiment.elec}),a.default.createElement("div",{className:"undertext"},"Fig.2: Electrophoretogram of our parts which we had submitted to registry, all of them show properly length (Using VF2 and VR primer, the length will increase about 300 bp)")),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.experiment.table_1}),a.default.createElement("div",{className:"undertext"},"Table 1: This table’s genes correspond to each lane of Fig. 2 from left to right.")))),c=a.default.createElement("div",null,"We use TCBS (Thiosulfate-citrate-bile salts-sucrose agar, a type of selective agar culture plate that is used in microbiology laboratories to isolate Vibrio) to examine Vibrio’s concentration in fish farm. Our HP member and wet lab member visit the fish farm in Taiwan, Chiayi. We collected the water sample from different pool, and sent it back to the lab to culture the sample on TCBS.",a.default.createElement("div",{className:"illustration"},a.default.createElement("div",{className:"illust-image-wrapper",style:{width:"600px"}},a.default.createElement("img",{className:"illust-image",src:o.default.image.experiment.sample})),a.default.createElement("div",{className:"undertext"},"Fig.3 : Collection sample from traditional fish farm in Taiwan")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Result"),a.default.createElement("div",null,"Sample from fish farm shown positive result, we found there were colony appeared on TCBS plate. Implying that Vibrio indeed exist in fish farm, and may infect shrimp or other marine creatures. In addition, the colony color stands for different types of Vibrio:"),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",null,"Vibrio Cholerae: Large yellow colonies. "),a.default.createElement("div",null,"Vibrio harveyi / Vibrio fischeri: Greyish-green to bluish-green colonies which show luminescence in dark.")),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:o.default.image.experiment.vibrio}),a.default.createElement("div",{className:"undertext"},"Fig.4 : Testing Sample from fish farm on TCBS"),a.default.createElement("div",{className:"undertext"},"Right plate shows Vibrio harveyi and the left pate shows Vibrio Cholerae")))),f=a.default.createElement("div",null,"Our detection system is designed to make E. coli to obtain the ability to sense the concentration of Vibrio in the environment. We found that most of the pathogenic Vibrio use “quorum sensing” to communicate with each individual and to express different genes. Quorum sensing, a system which is able to control the gene expression followed by the population density. The signal transmission of quorum sensing is totally depending on the auto-inducer, acyl homoserine lactones (AHL), a small chemical molecule which is produced by bacterial enzyme (Fig. 5). Moreover, the concentration of AHL is directly proportional to bacteria density. When AHL concentration exceeds the threshold, it will bind with the protein (LuxR) and trigger the promoter (pLUX) to express the downstream genes.",a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:o.default.image.experiment.ahl}),a.default.createElement("div",{className:"undertext"},"Fig. 5: The schematic of our gene circuits")),a.default.createElement("div",{className:"illustration",style:{width:"300px"}},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:o.default.image.experiment.structure}),a.default.createElement("div",{className:"undertext"},"Fig. 6: The structure of AHL")),"Our detection system is using Lux system. When the Vibrio density is high, the AHL concentration in the environment is high. Hence, our engineered E. coli can detect the Vibrio density and can release the anti-Vibrio substances. (Fig. 7)",a.default.createElement("div",{className:"illustration"},a.default.createElement("div",{className:"illust-image-wrapper",style:{width:"500px"}},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:o.default.image.experiment.cycle})),a.default.createElement("div",{className:"undertext"},"Fig. 7: The schematic of our detection system")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Exp. 1: AHL Sensor with Quantitative AHL "),a.default.createElement("div",null,"We constructed our AHL detector (BBa_K2774000), which can produce LuxR consistently by pLac promoter and can express GFP while LuxR combine with AHL and trigger pLUX (R0062). While AHL presents, AHL and LuxR will form a complex and trigger pLUX to express GFP. (Fig. 8)"),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.experiment.sensor}),a.default.createElement("div",{className:"undertext"},"Fig. 8: AHL Bio-Detector")),a.default.createElement("div",null,"In order to test our gene’s function, we added different concentration of AHL (3-oxo-C6-HSL, synthesized by Sigma) to DH5α cells transformed with BBa_K2774000, and measured GFP expression hourly."),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Procedure:"),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",{className:"innerpart"},"1. Prepare DH5α cells which transformed with BBa_K2774000 and liquid culture in LB overnight."),a.default.createElement("div",{className:"innerpart"},"2. Diluted eight bacterial culture samples in fresh LB (50 μL O/N culture with 4950 μL fresh LB)"),a.default.createElement("div",{className:"innerpart"},"3. Pre-culture until O.D.600 reached 0.4, making sure bacteria is growing in log phase (better expression capability) and add IPTG (1mM) to induce the expression of LuxR."),a.default.createElement("div",{className:"innerpart"},"4. Add different amount of AHL into different samples (0.01, 0.1, 0.5, 1, 5, 10, 100 nM) and 0 M AHL for positive control."),a.default.createElement("div",{className:"innerpart"},"5. Measure the O.D.600 value and expression of GFP (λexcitation = 490 nm; λemission = 530 nm) hourly.")),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.experiment.exp_1}),a.default.createElement("div",{className:"undertext"},"Fig. 9: GFP expression from different conc. of AHL"))),a.default.createElement("div",{className:"part"}),a.default.createElement("div",{className:"part-title"},"Result:"),a.default.createElement("div",null,"As the chart shows, we can find that AHL induced GFP expression as we expected. As the AHL concentration increased, the GFP expression increased until saturation. Besides, the optimized AHL concentration of inducing BBa_K2774000 is about 0.5-10 nM. Moreover, we found that if the concentration of AHL exceeds 1000 nM, it will inhibit BBa_K2774000 GFP expression."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:o.default.image.experiment.data_1}),a.default.createElement("div",{className:"undertext"},"Fig. 10: GFP expression from different conc. of AHL"))),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Exp. 2 : AHL sensor co-culture with fake Vibrio test"),"Since we cannot conduct the experiment related to Vibrio in our lab, we have to construct a fake Vibrio (by our engineered E. coli), which can also secrete AHL as Vibrio does. Therefore, we created our “pLac AHL generator” (BBa_2774004) by assembling “pLac + B0034” (BBa_J04500) and “LuxI” (BBa_C0161) together. Moreover, we added RFP to detect the population of fake vibrio (BBa_2774005)",a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Procedure:"),a.default.createElement("div",{className:"innerpart"}," 1. Prepare DH5α cells which transformed with BBa_2774000 (AHL sensor), BBa_2774004 (pLac AHL generator), and BBa_2774005 (pLac AHL generator + RFP) and liquid culture in LB overnight."),a.default.createElement("div",{className:"innerpart"}," 2. Diluted eight overnight bacterial culture in fresh LB with 1mM IPTG (50μL O/N culture with 4950 μL fresh LB)."),a.default.createElement("div",{className:"innerpart"}," 3. Pre-culture until O.D.600 reaches 0.4, making sure bacteria is growing in log phase (better expression capability)."),a.default.createElement("div",{className:"innerpart"}," 4. Mix different volume ratio of:"),a.default.createElement("div",null," BBa_2774004 (pLac AHL generator) with BBa_2774000 (AHL sensor)    "),a.default.createElement("div",null," BBa_2774005 (pLac AHL generator + RFP) with BBa_2774000 (AHL sensor)"),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:o.default.image.experiment.table_2})),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:o.default.image.experiment.table_3})),a.default.createElement("div",null,"5. Measure O.D.600, GFP and RFP expression after overnight culture.")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Result:"),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"a. BBa_2774005 (pLac AHL generator + RFP) co-culturing with BBa_2774000 (AHL sensor)"),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:o.default.image.experiment.exp_2}),a.default.createElement("div",{className:"undertext"},"Fig 11. AHL sensor with increasing concentration of AHL generator + RFP (from the left tube to the right tube). We centrifuged culture tubes in the left picture to get the ones in the right picture.")),a.default.createElement("div",{className:"innerpart"},"We co-cultured AHL sensor with increasing concentration of AHL generator + RFP. The picture indicates that AHL was indeed generated. When the two bacteria were mixed together, red and green fluorescence were both emitted and producing different colors.")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"b. BBa_2774004 (pLac AHL generator) co-culturing with BBa_2774000 (AHL sensor) "),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:o.default.image.experiment.table_4})),a.default.createElement("div",{className:"innerpart"},"In this experiment, we co-cultured AHL sensor with increasing concentration of AHL generator. We discovered that all liquid culture emitted green fluorescence except the one without adding AHL generator. We successfully proved that BBa_2774004 can express LuxI (AHL synthase) and generate AHL. We proposed that AHL concentration is linearly proportional to fake Vibrio concentration. However, the fluorescence intensities were almost the same among samples with increasing concentrations of AHL. We failed to construct a rational relation between fake Vibrio density and AHL concentration. We inferred the reason for this observation is that the rate of AHL synthesis is much higher than the rate of AHL degradation. The LuxI we used was the one without LVA-tag, so LuxI didn’t degrade instantly. In addition, we left the liquid culture overnight before measuring its fluorescence. Consequently, AHL was over-generated and the concentration of AHL was beyond the detection limit of AHL sensor.")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Exp. 3: Culture AHL sensor with Fake Vibrio’s supernatant")),a.default.createElement("div",null,"During Exp. 2 we find that overnight culturing and strong promoter cause excess AHL, which will not show the evidence of AHL concentration proportion to fake Vibrio concentration. Therefore we try a different way to experiment, we use weaker promoter and measure AHL concentration by AHL bio-sensor every hourly. Using interlab data we can get bacteria concentration by measuring OD600, and meanwhile we can centrifuge the tube to get supernatant which contain AHL secrete by the AHL generator that time."),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Procedure:"),a.default.createElement("div",{className:"innerpart"},"1. Prepare DH5α cells which transformed with BBa_2774000 (AHL sensor), BBa_2774002 (J23119+LxuI), and liquid culture in LB overnight."),a.default.createElement("div",{className:"innerpart"},"2.  Culture BBa_2774002 at microcentrifuge tube with (50μL O/N culture with 4950 μL fresh LB)."),a.default.createElement("div",{className:"innerpart"},"3. Measure microcentrifuge’s OD600 two hour and meanwhile centrifuge the tube to get supernatant contain AHL. Transfer the OD600 data to bacteria density by using inter lab data."),a.default.createElement("div",{className:"innerpart"},"4. Move 2.5 ml of supernatant contain AHL and 2.5ml fresh LB in new tube, and add AHL sensor simultaneously."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:o.default.image.experiment.exp_3}),a.default.createElement("div",{className:"undertext"},"Fig 12. The procedure of experiment 3."))),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Result:"),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:o.default.image.experiment.result_4}),a.default.createElement("div",{className:"undertext"},"Fig 13. AHL sensor with increasing concentration of AHL generator supernant")),a.default.createElement("div",{className:"innerpart"},"We transferred the supernatant from generator liquid culture which contained AHL into AHL sensor. The result showed that supernatant from a higher concentration of generator inducing AHL sensor to express higher amount of GFP. Moreover, after we transform the data and take the logarithm of generator density, we find a similar result which we had done in experiment 1. (Fig10)"),a.default.createElement("div",{className:"innerpart"},"Comparing exp1 and exp3 result, confirming that Vibrio density is proportional to the AHL concentration, at the condition of the real-time situation to avoid AHL accumulation. ")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Conclusion:"),a.default.createElement("div",null,"We have successfully created a bio-sensor which can detect and react to different concentration of AHL. Moreover, our fake Vibrio (BBa_2774004) can also secrete AHL. We also confirmed that the Vibrio density is proportional to the AHL concentration. Our future work is to exchange gene which is under the control of pLux from GFP to anti-Vibrio substances."))))),d=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"innerpart"},"Based on our gene circuit, we need two kinds of substances to fulfill our goal. The first one is the suicide gene. When Vibrio concentration is low, our E. coli need to suicide themselves to maintain the equilibrium. Therefore, we introduced “tsRNA” (toxic small RNA), which was studied and experimented by Dr. Rotem . “tsRNA” was first found in Burkholderia cenocepacia and showed its toxic to E. coli. to inhibit E. coli growth. Moreover, references show that “tsRNA” couldn’t penetrate the bacterial cell wall, either from outside or inside. Therefore, it becomes the best candidate for our suicide gene. Although the mechanism of “tsRNA” is not clearly known, Dr. Rotem had tried complement sequences of “tsRNA” shown negative results. They hypothesized that “tsRNA” might target and mis-regulate essential genes within the E. coli cell. "),a.default.createElement("div",{className:"innerpart"},"To test the function of tsRNA for our suicide gene, we put tsRNA sequence under pLac promoter. Using IDT synthesized technology, we got the gene fragment (BBa_2774001)."),a.default.createElement("div",{style:{width:"300px"},className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:o.default.image.experiment.tsrna}),a.default.createElement("div",{className:"undertext"},"Fig 14. The secondary structure of tsRNA"))),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Procedure"),a.default.createElement("div",{className:"innerpart"},"1. Prepare DH5α which transformed with BBa_K2774001 and liquid culture in LB overnight."),a.default.createElement("div",{className:"innerpart"},"2. Diluted the overnight bacterial culture in fresh LB (50 μL O/N culture with 4950 μL fresh LB) "),a.default.createElement("div",{className:"innerpart"},"3. Pre-culture until O.D.600 reaches 0.2, making sure bacteria is growing in log phase (better expression capability) "),a.default.createElement("div",{className:"innerpart"},"4. Add IPTG (1mM) to induce tsRNA expression. (Adding ampicillin for positive control)"),a.default.createElement("div",{className:"innerpart"},"5. Measure O.D.600 hourly."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:o.default.image.experiment.tsrna_test}))),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Result"),a.default.createElement("div",{className:"innerpart"},"We induced tsRNA expression with IPTG at O.D.600 = 0.2. The result shows that tsRNA slightly inhibits E. coli growth, approximately 12%. We tested our BBa_K2774001 lots of times, but it still goes to the same result. Feeling confused, so we consulted Dr. Rotem for our situation, and got a generous reply:"),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:o.default.image.experiment.email})),a.default.createElement("div",{className:"innerpart"},"Dr. Rotem suggested us to check if our tsRNA mutated during transforming because of the inferiority of growth rate. Or to check tsRNA expression by using Northern blotting to check the RNA expression. Therefore, we sequenced our plasmid (tsRNA) and BLAST the result with our designed sequence: showing no mutation on tsRNA section."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:o.default.image.experiment.strength}),a.default.createElement("div",{className:"undertext"},"Fig 15. Sequencing result of tsRNA")),a.default.createElement("div",{className:"part-title"},"Conclusion"),a.default.createElement("div",{className:"innerpart"},"Our experiment result shows that the inhibition efficiency of tsRNA is 12%, which is significantly different from the one reported by Dr. Rotem. We sequenced our plasmid and no mutation was found. We inferred that the low inhibition efficiency is due to few expression of tsRNA. We believe this can be improved by using a stronger promoter to get a better result."))),p={viewbar:{Title:"Experiment",Subtitle:"",image:o.default.image.topfull.experiment},content:{Introduction:"In order to achieve our project’s goal, we constructed a few functional genes and tested their features. To sum up, our project can be divided into three parts. First of all is the “detection” part, we made our E. coli to detect the density of Vibrio and to respond. Then, it’s the part of “kill Switch”, we made our E. coli to switch from secreting anti-Vibrio substances to suicide to fulfilled bio-safety issues. Finally, it’s  “substances test” for anti-Vibrio and suicide substances which will be used in our project.",parts:[{Title:"Cloning",Description:u},{Title:"Testing Sample",Description:c},{Title:"Detection",Description:f},{Title:"Substances test",Description:d}]}},h=function(e){function t(e){return function(e,t){if(!(e instanceof t))throw new TypeError("Cannot call a class as a function")}(this,t),function(e,t){if(!e)throw new ReferenceError("this hasn't been initialised - super() hasn't been called");return!t||"object"!=typeof t&&"function"!=typeof t?e:t}(this,(t.__proto__||Object.getPrototypeOf(t)).call(this,e))}return function(e,t){if("function"!=typeof t&&null!==t)throw new TypeError("Super expression must either be null or a function, not "+typeof t);e.prototype=Object.create(t&&t.prototype,{constructor:{value:e,enumerable:!1,writable:!0,configurable:!0}}),t&&(Object.setPrototypeOf?Object.setPrototypeOf(e,t):e.__proto__=t)}(t,i.Component),r(t,[{key:"render",value:function(){return 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We both aimed to let students know more about iGEM and synthetic biology. During the camp, participants learned how to carry out simple experiments, e.g. plasmid extraction and transformation. We also shared with the students about our iGEM project and briefly introduced synthetic biology. The following are the schedule and all the material we used in the camp. About our files used in this camp are all available to",a.default.createElement("a",{href:"https://drive.google.com/drive/folders/19QhZWbbpJTddxGISAeXFJBAxcs_MuS_m?usp=sharing"}," download"),". The only thing you have to notice is the language, most are in Chinese."),f=a.default.createElement("div",null,a.default.createElement("div",{className:"illustration"},a.default.createElement("table",{className:"table"},a.default.createElement("tbody",null,a.default.createElement("tr",{className:"tablehead"},a.default.createElement("th",{className:"tablehead"}),a.default.createElement("th",{className:"tablehead"},"2018/08/10"),a.default.createElement("th",{className:"tablehead"},"2018/08/11")),a.default.createElement("tr",{className:"tablehead"},a.default.createElement("th",{className:"tablehead"},"9:00~12:00"),a.default.createElement("td",{className:"tabledata"},a.default.createElement("div",null,"iGEM Introduction & Team Project"),a.default.createElement("div",null,"Experiment Introduction "),a.default.createElement("div",null,"Pipetman Teaching ")),a.default.createElement("td",{className:"tabledata"},a.default.createElement("div",null,"Synthetic Biology Introduction"),a.default.createElement("div",null,"Experiment Introduction"),a.default.createElement("div",null,"Colony PCR"))),a.default.createElement("tr",{className:"tablehead"},a.default.createElement("th",{className:"tablehead"},"12:00~13:00"),a.default.createElement("td",{className:"tabledata"},"Lunch Time"),a.default.createElement("td",{className:"tabledata"},"Lunch Time")),a.default.createElement("tr",{className:"tablehead"},a.default.createElement("th",{className:"tablehead"},"13:00~16:00"),a.default.createElement("td",{className:"tabledata"},a.default.createElement("div",null,"Plasmid Extraction"),a.default.createElement("div",null,"Transformation"),a.default.createElement("div",null,"Bacteria Cultivation")),a.default.createElement("td",{className:"tabledata"},a.default.createElement("div",null,"Colony PCR"),a.default.createElement("div",null,"DNA Electrophoresis"),a.default.createElement("div",null,"Questionnaire Survey"))))))),d=a.default.createElement("div",null,a.default.createElement("div",null,"This year, 16 students joined our synthetic biology camp. The following are some feedback given by the students in the questionnaire survey. 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Or the students would be boring."),a.default.createElement("tr",{className:"tablehead"},a.default.createElement("td",{className:"tabledata"},"The contents of the experiments were quite funny, but the basic theories were difficult for high school students.")),a.default.createElement("tr",{className:"tablehead"},a.default.createElement("td",{className:"tabledata"},"Though I have not learned most of the terms about molecular biology experiments, the contents of experiments were interesting and inspiring which makes me acquire knowledge about molecular biology.")),a.default.createElement("tr",{className:"tablehead"},a.default.createElement("td",{className:"tabledata"},"If lecturers summarized all of the experiments to a table, it will be better!")),a.default.createElement("tr",{className:"tablehead"},a.default.createElement("td",{className:"tabledata"},"It’s my first time to do the molecular biology experiments and hopefully, you guys can achieve your goals!"))))))),p=a.default.createElement("div",null,a.default.createElement("div",null,"According to their feedback, we have achieved our goal: First, to introduce iGEM and synthetic biology. Second, teach high school students how to carry out synthetic biology experiment. Third, enhance their interest in genetic engineering. The synthetic biology camp this year is successful, and National Feng-Shan Senior High School are willing to cooperate with us next year! We hope our effort can encourage more students to do researches about genetic engineering, and maybe they will join iGEM soon!"),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:l.default.image.education.group}))),h={viewbar:{Title:"Education",Subtitle:u,image:l.default.image.topfull.education},content:{Introduction:"Joining iGEM allows us to carry out some interesting and creative experiments related to the latest researches. However, for ordinary senior high school students in Taiwan, they do not have the opportunity to engage in iGEM or the state-of-the-art knowledge of synthetic biology. Therefore, we decided to bring synthetic biology to high school students. We can not only tell them what synthetic biology is but also share our experience of joining iGEM as well as encourage them to participate next year!",parts:[{Title:"Summer Synthetic Biology Camp",Description:c},{Title:"Schedule",Description:f},{Title:"Feedback",Description:d},{Title:"Summary",Description:p}]}},m=function(e){function t(e){return function(e,t){if(!(e instanceof t))throw new TypeError("Cannot call a class as a function")}(this,t),function(e,t){if(!e)throw new ReferenceError("this hasn't been initialised - super() hasn't been called");return!t||"object"!=typeof t&&"function"!=typeof t?e:t}(this,(t.__proto__||Object.getPrototypeOf(t)).call(this,e))}return function(e,t){if("function"!=typeof t&&null!==t)throw new TypeError("Super expression must either be null or a function, not "+typeof t);e.prototype=Object.create(t&&t.prototype,{constructor:{value:e,enumerable:!1,writable:!0,configurable:!0}}),t&&(Object.setPrototypeOf?Object.setPrototypeOf(e,t):e.__proto__=t)}(t,i.Component),r(t,[{key:"render",value:function(){return a.default.createElement(o.default,h)}}]),t}();t.default=m},function(e,t,n){"use strict";Object.defineProperty(t,"__esModule",{value:!0});var r=function(){function e(e,t){for(var n=0;n<t.length;n++){var r=t[n];r.enumerable=r.enumerable||!1,r.configurable=!0,"value"in r&&(r.writable=!0),Object.defineProperty(e,r.key,r)}}return function(t,n,r){return n&&e(t.prototype,n),r&&e(t,r),t}}(),i=n(0),a=s(i),o=(s(n(10)),s(n(4))),l=s(n(8));function s(e){return e&&e.__esModule?e:{default:e}}var u=a.default.createElement("div",null,a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.interview.research})),a.default.createElement("div",null,"Fisheries Research Institute, COA, which is the government agency offers fish farmers to test the pathogen of fish and also, doing some researches on aquatic organisms as well. Deputy director of COA, C.I. Chang, who is a master in marine biology, recently did lots of researches on aquatic animal and pathogen, creating the MV-kit as well. The kit which function is to test the number of Vibrio in the water. And of course, FRI is the closest department between the local fish farmers and the public. As a result, we decided to visit him to get more helpful information and suggestions for our project. During our in-depth talk, we knew more about the Vibrio and, our core of the automated device, ACE (link to drylab ACE design). For example, after knowing the detailed information about the basic characteristics of Vibrio and the mechanism of MV-kit, making us want to combine the kit with our device and if it comes true, the device will be powerful. In the end, deputy director also gave us a few useful, potential advice and questions between our device and the kit. These feedbacks inspired us pretty much, such as whether our device would be influenced by the precipitation or not, or do we need some kinds of membrane to filter the aquatic sample or not, and in order to maintain the integrity and accuracy of our device, we have to test the efficiency of MV-kit in different environments and to find the simplest way that fish farmers can easily use. Nicely, the deputy director offering the plate of Vibrio harveyi and MV-kit for free in order to encourage us to make our experiment more complete.")),c=a.default.createElement("div",null,"In order to improve our project, we need to gain as much feedback as possible from the potential users; therefore, we’ve visited local fisherman in Taiwan, of which we can categorize into ecological and traditional breeding fish farmers."),f=a.default.createElement("div",null,a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.interview.tradition_1})),a.default.createElement("div",null,a.default.createElement("div",null,"In addition to visiting the ecological breeding farmers, we visited traditional fishermen in Kaohsiung and Chiayi, Mr. Chiu and Mr. Jian. They gave us some useful information, so we can improve our project more further."),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Mr. Chiu and Mr. Jian"),a.default.createElement("div",{className:"innerpart"},"Overall, they usually wait until the fish get sick and take them to the inspection center. If we can let them know about the condition of bacteria beforehand, they might take precautions, such as cleaning the water or adding the active powder to improve water quality and minimize the damage."),a.default.createElement("div",{className:"innerpart"},"When asking about what problem is the most crucial one they are facing, they’ve mentioned the uncertainty of electricity.  If the machine shut down or in a dangerous condition, sending a notification to farmers is essential. They also mentioned that adding the function of notifying farmer by message might be more practical than the smartphone application or the notification bar since farmer might be the elderly who do not use a smartphone. In light of their suggestions, we found the Chunghwa Telecom to seek for future collaboration. They told us that as long as we apply for a SIM card and a phone number, this kind of function can easily be done."),a.default.createElement("div",{className:"innerpart"},"To sum up, feedbacks and information are listed below. Plenty of them has greatly improved our projects:"),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",null,"1. Their working time is usually between 2 a.m. to 11 a.m., so they suggest us to test the sample during this time (since MV-kit needs 12~24 hr. to react completely)."),a.default.createElement("div",null,"2. They introduce the “TCBS” method: a common way to detect Vibrio sp. in traditional fish farm. This method need 1~2 day to complete."),a.default.createElement("div",null,"3. Add more features to ACE machine, e.g. water quality test (pH, NO3-, NO2-), other kit test (virus, fish parasite), and black out reminder."),a.default.createElement("div",null,"4. They recommend us to use text messages to be the notification of ACE machine. Because some of the traditional fishermen do not use smartphones."),a.default.createElement("div",null,"5. They recommend us to cooperate with fishermen who feed grouper or eel."),a.default.createElement("div",null,"6. As a traditional fisherman, they think our device has great potential."))))),d=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Mr. Tsai Hung-Jing’s Fish Farm"),a.default.createElement("div",null,a.default.createElement("div",{className:"innerpart"},"Before deciding which fish farm to visit, we looked for information on the Internet and finally found an interview published by CommonWealth Magazine, which is a famous magazine in Taiwan. The article showed that how successfully Mr. Tsai uses biological feed - water flea to take care of his shrimps and to make some profit. This idea of substituting chemical feed into water flea, an eco-friendlier one, matched the concept of iGEM. So, we called him and arranged the visit time without hesitation."),a.default.createElement("div",{className:"innerpart"},"Mr. Tsai was nice and friendly, he not only introduced his fish farm in such a detailed way but also answered all of our questions. He mentioned that the way he used to prevent Vibrio is to make the environment beneficial for good germs, which occupies the environment of the Vibrio, leaving no spaces for them to grow. He pointed out that people often uses ClO2 to kill Vibrio is unable to eradicate the problem completely. Once the pollutants are in, Vibrio will grow again. Consequently, we introduced our method to him and he sounded satisfied and looked forward to our final result. He also gave us some positive feedback toward our ACE. He said that if our service cost can be as low as 8,000 NT dollars, it will have a strong advantage and the market acceptance will be high since the price of the general detection devices need to be up to ten thousand NT dollars."),a.default.createElement("div",{className:"innerpart"},"The interview with Mr. Tsai really cheered us up, he let us understand the thing we do might help them in a practical way. Even though we went back to school, he still sent messages to encourage us.")),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.interview.eco_1}))),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Qiujia Aquaculture in Chiayi County"),a.default.createElement("div",null,a.default.createElement("div",{className:"innerpart"},"Qiujia Aquaculture has been established for 50 years, the main breed they sell are milkfish and shrimps. The reason why we wanted to visit them was that they refuse to use any of the chemical agents or drug and it triggered our curiosity, how do they make it?"),a.default.createElement("div",{className:"innerpart"},"By actually visiting them on the ground, we discovered that they do not use any chemical medicine or drug to kill bacteria, instead, they use Bacillus subtilis to compete with Vibrio. We surprisingly found out this kind of ecological competence is similar to what our wet lab is trying to mimic. Though adding good germs can’t guarantee no fish will be infected, they believed doing so was far better than adding artificial chemicals which have huge uncertainty to humankind. When asking about whether they would detect the concentration of Vibrio, they told us the current method they use to detect Vibrio is coating the plate, which is not efficient and not so accurate."),a.default.createElement("div",{className:"innerpart"},"Furthermore, they provided us with pieces of advice and feedback regarding our Automatic Color-Detecting Examiner (ACE):"),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",null,"1.ACE needs to be equipped with high accuracy, shutdown notification, and self-calibration."),a.default.createElement("div",null,"2. If they knew the concentration of Vibrio earlier, they can take actions to prevent the Vibrio from growing too much beforehand, such as adding Bacillus subtilis."),a.default.createElement("div",null,"3. Since the growing time of Vibrio to be pathogenic is way longer than one day, a period of sampling doesn’t need to be as frequent as 1 hour, but one day. "),a.default.createElement("div",null,"4. They suggested us to target indoor breeding customers, since that if the device is set outdoors, it may be easily broken down. Moreover, fish with high economic value bred indoor might be more fragile than the outdoors.")),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.interview.eco_2})),a.default.createElement("div",null,"After considering their suggestions, we decided to extend the sampling period to 12 hours, adding the fool-proof design, such as that if the camera shut down or cannot be connected, it will return a “no camera” sign to our application.")))),p=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},a.default.createElement("div",null,a.default.createElement("div",{className:"part-title"},a.default.createElement("a",{href:"http://dataweb.nmmba.gov.tw/ResearchMemberDetail.aspx?CondID=bfcdc17d-da5f-48f8-9af8-819a3376101a"},"Dr. Jimmy Kuo")),"Dr. Jimmy Guo, a vice researcher of the national museum of marine biology and aquarium, have done many types of research of Vibrio in the aquarium. He first shared with us how he made the research on Vibrio and explained why the growth curve of Vibrio was so strange. The reason is that Vibrio is a special species which is different from other normal bacteria. Vibrio will be dormant and make its metabolism disordered when stored at low temperature. Therefore, if we want to get the expectative results, we should culture fresh Vibrio rather than take one from the refrigerator."),a.default.createElement("div",null,"Besides the advice for the wet lab, Dr. Guo also gave us some suggestions to our hardware. He recommended that we could use the battery as our electric power source and thus, avoiding the loss of results from the blackout. Additionally, our device should calibrate regularly, or the results would be deviant. Finally, since our device could detect frequently, we could target on the customers cultivating remunerative aquatic agriculture."),a.default.createElement("div",null,a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.interview.aqua_1})))),a.default.createElement("div",{className:"part"},a.default.createElement("div",null,a.default.createElement("div",{className:"part-title"},a.default.createElement("a",{href:"https://www.nmmba.gov.tw/research/RMenber_Content.aspx?n=291A05EEDDA1D4C4&sms=CF7FFA69EB91FF61&s=6A6B57F5FE966020"},"Dr. Ming-An Tsai ")),"Dr. Ming-An Tsai, an assistant researcher in the national museum of marine biology and aquarium, is interested in doing research on diseases of aquatic animals. Fortunately, Dr. Tsai’s lab is also doing some Vibrio-detecting experiment, so we first exchanged some ideas about Vibrio experiments. After introducing our project, Dr. Tsai gave us some important advice. First of all, he suggested us that we should ensure which way we would use to avoid bacterial diseases, killing Vibrio or inhibiting its growth. Additionally, he reminded us that we should not kill all the Vibrio in the water, or we might destroy the ecological balance. Finally, he gave us a crucial reference that if the concentration of Vibrio in the water goes beyond 104 CFU/mL, it would be more infectious. This data is a vital reference not only in our wet lab but also in our dry lab."),a.default.createElement("div",null,a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.interview.aqua_2}))))),h={viewbar:{Title:"Interview",Subtitle:"Visited the research institute,fishing farm, and aquarium",image:o.default.image.topfull.interview},content:{Introduction:"Our project is an interdisciplinary work which is so comprehensive and complete to makes it possesses a potent potential to practice in the real world. Based on this advantage, to navigate our project toward a suitable direction to either meet the true demand and solve real problem is quite essential. At the beginning, we visited Deputy director of Fisheries Research Institute. He gave us some advice on how to start our project. Then, we had conducted interviews with fishermen in Taiwan, visited the fishing farm with them and received countless useful information about how to alter our design to fit their demand and improve the latest solution they adopt to combat bacterial infection. Moreover, with the advice given by researchers from national museum of marine biology and aquarium, we gained much more practical perspective on what’s the pros and cons of our solution and how we can lower the risk that will harm the environment. With their help, we have achieved the core value of human practice: know who need, find what they need and meet their need.        ",parts:[{Title:"Fisheries Research Institute, COA ",Description:u},{Title:"Interview with Fish Farmer",Description:c},{Title:"Ecological Breeding",Description:d},{Title:"Traditional Fishermen",Description:f},{Title:"National Museum of Marine Biology and Aquarium",Description:p}]}},m=function(e){function t(){return function(e,t){if(!(e instanceof t))throw new TypeError("Cannot call a class as a function")}(this,t),function(e,t){if(!e)throw new ReferenceError("this hasn't been initialised - super() hasn't been called");return!t||"object"!=typeof t&&"function"!=typeof t?e:t}(this,(t.__proto__||Object.getPrototypeOf(t)).apply(this,arguments))}return function(e,t){if("function"!=typeof t&&null!==t)throw new TypeError("Super expression must either be null or a function, not "+typeof t);e.prototype=Object.create(t&&t.prototype,{constructor:{value:e,enumerable:!1,writable:!0,configurable:!0}}),t&&(Object.setPrototypeOf?Object.setPrototypeOf(e,t):e.__proto__=t)}(t,i.Component),r(t,[{key:"render",value:function(){return a.default.createElement(l.default,h)}}]),t}();t.default=m},function(e,t,n){"use strict";Object.defineProperty(t,"__esModule",{value:!0}),t.default=void 0;var r=function(){function e(e,t){for(var n=0;n<t.length;n++){var r=t[n];r.enumerable=r.enumerable||!1,r.configurable=!0,"value"in r&&(r.writable=!0),Object.defineProperty(e,r.key,r)}}return function(t,n,r){return n&&e(t.prototype,n),r&&e(t,r),t}}(),i=n(0),a=s(i),o=(s(n(10)),s(n(4))),l=s(n(8));function s(e){return e&&e.__esModule?e:{default:e}}var u=a.default.createElement("div",null,a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.public.nctu})),a.default.createElement("div",null,"Engaging with other iGEM teams to exchange ideas and to brainstorm together is a crucial part of iGEM. Therefore, we joined a half-day meet-up with three iGEM teams in Taiwan, NTHU_Formosa, NCTU_Formosa, and Mingdao. During the meet-up, we presented our projects, had some discussions with other teams, and received some problems and feedbacks on our projects. After brainstorming, we knew what the problem was and what we could do to improve the project. This meet-up truly broadened our horizons and we got loads out of it. Following are some feedbacks we received from the meet-up:"),a.default.createElement("div",null,a.default.createElement("div",{className:"part"}," 1. There were some drawbacks of our project of biosafety if we put our engineered E. coli into the open water area. After discussion, we finally decided to replace this way by putting it into a close water and to use it with a filter. By doing so, we could effectively control our engineered bacteria and thus, reducing the problem about biosafety. "),a.default.createElement("div",{className:"part"}," 2. We should ensure our device can really benefit the users. Therefore, we visited some local fishermen in person and sought for their suggestions. To find out more, check the ",a.default.createElement("a",{href:"https://2018.igem.org/Team:NTHU_Taiwan/Interview"},"interview")," page"))),c=a.default.createElement("div",null,a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.public.meetup_group})),a.default.createElement("div",null,"In late July, we participated in the first Eurasian meet-up held by BGI college and TU Delft in Shenzhen, China. There are 20 stunning teams from 5 different countries visited Shenzhen, discussing and sharing all kinds of iGEM issues and projects during the conference. Also, we presented and conveyed our idea to other teams, which lets us receive several feedbacks and they were quite useful for our project. At this point, we did find some inspiring ideas which sharpen our current work. Here we conclude something that influenced our work:"),a.default.createElement("div",null,a.default.createElement("div",{className:"part"},"1. The other iGEMers provided us with some useful feedbacks, for instance, they suggested us to integrate and double-check the consistency of data from our bio-reactor and Vibrio-detecting system. Also, team members from CUHK gave us some suggestions about our image detection system. Moreover, some people suggested us other switch systems for our experimental design like toehold switch. After the conference, we searched for several switch systems but we still stick to our original idea because we thought that the disadvantage for our STAR system is the least."),a.default.createElement("div",{className:"part"},"2. A more concise presentation should be considered. This is the first presentation after our frame was formed out, but we spent much time explaining the detailed information about our project. Furthermore, we didn’t integrate our wet lab works and dry lab works in our presentation which was not convincing to the audience. After the presentation, we then modified the whole slides into a more concise and more comprehensible version which do help more audiences to get into our story."),a.default.createElement("div",{className:"part"},"3. The meet-up official set up several workshops and lectures during the conference. For instance, they invited former iGEM judge and iGEM ambassador to share their experiences like “how to be an iGEMer?” from many different perspectives. It seems to us that we learned more details and tips about how to handle the iGEM project and figured out the overview of this competition.")),a.default.createElement("div",null,"Last but not least, except for the feedback we received, we also build a strong friendship connection there. The meet-up official assigns a buddy team for each team and our buddy is Marburg university from Germany. We really enjoyed the moment with them. "),a.default.createElement("div",null,a.default.createElement("div",{className:"gallery"},a.default.createElement("img",{style:{width:"45%",margin:"0px 1%"},src:o.default.image.public.meetup_self}),a.default.createElement("img",{style:{width:"45%",margin:"0px 1%"},src:o.default.image.public.meetup_presentation})))),f=a.default.createElement("div",null,a.default.createElement("div",{className:"illustration"},a.default.createElement("iframe",{style:{height:"500px",width:"100%"},src:"https://www.youtube.com/embed/1wib7JQEZ78"})),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"About TRANS"),a.default.createElement("div",null,"TRANS Conference is a platform where entrepreneurs, experts, investors, and students can exchange ideas, which was created by H. Spectrum, a startup incubator/accelerator focusing on the healthcare sector. As we had a chance to join in this forum, we decided to present our project to people in the healthcare field as many as possible, hoping to get some feedback from them and improve our project."),a.default.createElement("div",null,a.default.createElement("div",{className:"gallery"},a.default.createElement("img",{style:{width:"45%",margin:"0px 1%"},src:o.default.image.public.hspectre}),a.default.createElement("img",{style:{width:"45%",margin:"0px 1%"},src:o.default.image.public.trans})))),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Discussion and feedback"),a.default.createElement("div",null,"During TRANS, we found that people who are interested in our project are from multiple different fields and not just professionals. We’ve met students, bacterial professionals, fishery companies and some previous iGEM teams in Taiwan, whose backgrounds are medical, biology, microbiology, culture studies, medical engineering, and electrical engineering."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.public.discuss_1})),a.default.createElement("div",{className:"part"},a.default.createElement("div",null,"After the discussion, we sorted out the most common questions from our listeners as below:"),a.default.createElement("div",null,a.default.createElement("div",null,"1. What is the principle of MV-kit and what’s its difference between PCR analysis?"),a.default.createElement("div",null,"2. Biosafety such as being eaten by people and genetic mutation of E.coli."),a.default.createElement("div",null,"3. Will engineered genes exchange with bacterial organisms in the environment, causing genetic recombination problems because of the complex environment?"),a.default.createElement("div",null,"4. E.coli residual problems."),a.default.createElement("div",null,"5. Is engineered E.coli regulation acceptable? GMO products in Taiwan are still very opposed, how to persuade?"),a.default.createElement("div",null,"6. The growth and decline effect of E. coli will still have resistance problems in the future. Is there any solution?"))),a.default.createElement("div",{className:"part"},a.default.createElement("div",null,"And we also got some feedback and advice:"),a.default.createElement("div",null,a.default.createElement("div",null,"1. The mechanism of STAR/anti-STAR can also be applied to the sterilization of intensive care units."),a.default.createElement("div",null,"2. We could transform our target to a smaller range to simplify our problem, such as moving our environment from outdoor to indoor."),a.default.createElement("div",null,"3. In the future, if our growth and decline mechanism success, we could experiment our genes by intelligent shrimp culturing."),a.default.createElement("div",null,"4. To solve the problem of drug resistance, try adding multiple attack lines to E. coli."),a.default.createElement("div",null,"5. A company called ID Water which major in water quality testing suggested us to cooperate with them and tried to start an undertaking."))),a.default.createElement("div",{className:"gallery"},a.default.createElement("img",{style:{width:"30%",margin:"0px 1%"},src:o.default.image.public.discuss_2_1}),a.default.createElement("img",{style:{width:"30%",margin:"0px 1%"},src:o.default.image.public.discuss_2_2}),a.default.createElement("img",{style:{width:"30%",margin:"0px 1%"},src:o.default.image.public.discuss_2_3})),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"innerpart"},"After digesting the advice, we decided to modify our project in detail by meeting some of the professional companies, which we have saved the contacting information. First, we emailed to ID Water, asking for a chance to visit their company, hoping to have a chance to communicate some detecting methods. For example, we have trouble in analyzing with fluorescence in a bioreactor, which is part of their technique in water analyzing. ID Water thus told us that their cooperator, IGC-BIO, a healthcare company in Taiwan, is also researching in the method of interaction of bacteria, so we also plan to visit them someday."),a.default.createElement("div",{className:"innerpart"},"We could also meet Hin-Chung Wong professor in Soochow University, who is a profession in Vibrio field, for more detailed information about Vibrio to perfect our project. And we have checked again that our project doesn’t violate the iGEM rule. In the future, if we want to test our bacteria, LiveStrong Biomedical Technology promised us that they would provide a shrimp culturing sink for us to practice our experiment. Finally, thank for those who gave us lots of precious advice."),a.default.createElement("div",{className:"gallery"},a.default.createElement("img",{style:{width:"45%",margin:"0px 1%"},src:o.default.image.public.discuss_3_1}),a.default.createElement("img",{style:{width:"45%",margin:"0px 1%"},src:o.default.image.public.discuss_3_2}))))),d=a.default.createElement("div",null,a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.public.xgem})),a.default.createElement("div",null,"Interacting with people from different backgrounds is amazing and we do cherish every moment we engaged with other iGEM teams to exchange ideas as well as share techniques. For us, bringing the people across the world together to brainstorm about synthetic biology and how to make this world a better place is the magic of iGEM. Therefore, we established a platform on Slack named XGEM. On XGEM, teams can share information and ask questions immediately. So far, 18 teams from 14 different countries have joined XGEM. More about the teams joining XGEM:"),a.default.createElement("div",null,a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.public.chart_1})),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.public.chart_2})),a.default.createElement("div",{className:"illustration",style:{width:"70%"}},a.default.createElement("img",{className:"illust-image",src:o.default.image.public.chart_3}))),a.default.createElement("div",null,"Its really exciting to have 19 teams from all over the world to interact on XGEM. Some teams have posted their questionnaires related to their project or shared what they had done. We really appreciate your participation and can’t wait to meet these teams in person at Giant Jamboree!")),p={viewbar:{Title:"Public",Subtitle:"Meetups, conferences, and online platform",image:o.default.image.topfull.public},content:{Introducation:"Due to the fact that the synthetic biology is a brand-new field for the general public. Consequently, creating an enlarging division of knowledge between the general public and synthetic biology is necessary, and thus the public engagement is crucial for bridging the gap. The following are the activities we attended this year. We learned so much from the public, our friends and other iGEM teams. 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Therefore, our goal this year is to design a system which is able to dose appropriate and corrected aquacultural drugs according to the concentration of bacteria in the water as well as automatically detect the water condition which can let fishermen have a comprehensive understanding of whether the drug works or not.")),c=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},"Our project can be divided into two parts: one is to dose appropriate and corrected drug, and the other is to automatically detect the water quality and the concentration of specific bacteria. "),a.default.createElement("div",{className:"part"},"To implement the idea of appropriately dosing aquaculture drug, we design a system based on the model of succession. 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The system regulates the rate of killing ",a.default.createElement("span",{className:"special-name"},"Vibrio")," and ",a.default.createElement("span",{className:"special-name"},"E. coli")," itself in order to keep the population of ",a.default.createElement("span",{className:"special-name"},"Vibrio")," under the pathogenic level and maintain the balance between two bacteria at the same time. Therefore, we can control the ",a.default.createElement("span",{className:"special-name"},"Vibrio")," population and avoid fish and shrimp infection in a sustainable way. "),a.default.createElement("div",{className:"part"},"In the part of detecting water quality, to get precise concentration of specific bacteria in the water, we design an automatic and real-time detecting device. The current detection methods are time-consuming and furthermore, the water quality monitoring system on the market currently is able to detect physical and chemical indexes only. Therefore, we aim to create an IoT detecting device which can collect the water sample automatically and periodically. Then it is able to detect the concentration of specific bacteria with high specificity. For instance, it can detect the concentration of 100 CFU/mL of ",a.default.createElement("span",{className:"special-name"},"Vibrio")," within 24 hours[4,5], and informs fisherman regularly. It has the potential to reduce the financial loss of our aquaculture industry. ")),f=a.default.createElement("div",null,a.default.createElement("div",{className:"reference"},"1. Ching-Ying Deng, Meei-Mei Chen, Ju-Hsien Peng, Yu-Chiao Yang, Chih-Chien Chang, Hui-Ping Tsai 2014 The Investigation on the Infection of Francisellosis in Farmed Tilapia in Taoyuan and Hsinchu Areas. Journal of Taiwan Fisheries Research,22 (1): 81-91"),a.default.createElement("div",{className:"reference"},"2. https://news.un.org/en/audio/2014/05/588932, UN NEWS. 19 May 2014 "),a.default.createElement("div",{className:"reference"},"3. H-J Ju, M-Y Huang, H-C Liu, L-W Tseng, C-L Pan and C-I Chang, 2016 Screening Bacteria Antagonistic toward Aquatic Pathogenic Vibrio spp. from Marine Fishes and Shrimp. Journal of Taiwan Fisheries Research, 24 (1): 37-50"),a.default.createElement("div",{className:"reference"},"4. Chin-I Chan, et al., 2011, https://www.tfrin.gov.tw/dl.asp?fileName=17111695971.pdf"),a.default.createElement("div",{className:"reference"},"5. http://www.tfrin.gov.tw/friweb/frienews/enews0118/h1.html")),d={viewbar:{Title:"Description",Subtitle:"A solution to the problem of the abuse of antibiotics in aquaculture",image:o.default.image.topfull.description},content:{Introduction:"",parts:[{Title:"Motivation",Description:u},{Title:"Solution",Description:c},{Title:"Reference",Description:f}]}},p=function(e){function t(){return function(e,t){if(!(e instanceof t))throw new TypeError("Cannot call a class as a function")}(this,t),function(e,t){if(!e)throw new ReferenceError("this hasn't been initialised - super() hasn't been called");return!t||"object"!=typeof t&&"function"!=typeof t?e:t}(this,(t.__proto__||Object.getPrototypeOf(t)).apply(this,arguments))}return function(e,t){if("function"!=typeof t&&null!==t)throw new TypeError("Super expression must either be null or a function, not "+typeof t);e.prototype=Object.create(t&&t.prototype,{constructor:{value:e,enumerable:!1,writable:!0,configurable:!0}}),t&&(Object.setPrototypeOf?Object.setPrototypeOf(e,t):e.__proto__=t)}(t,i.Component),r(t,[{key:"render",value:function(){return a.default.createElement(l.default,d)}}]),t}();t.default=p},function(e,t,n){"use strict";Object.defineProperty(t,"__esModule",{value:!0}),t.default=void 0;var r=function(){function e(e,t){for(var n=0;n<t.length;n++){var r=t[n];r.enumerable=r.enumerable||!1,r.configurable=!0,"value"in r&&(r.writable=!0),Object.defineProperty(e,r.key,r)}}return function(t,n,r){return n&&e(t.prototype,n),r&&e(t,r),t}}(),i=n(0),a=s(i),o=s(n(4)),l=s(n(8));function s(e){return e&&e.__esModule?e:{default:e}}var u=a.default.createElement("div",null,a.default.createElement("div",null,"Our project this year is to design an engineered E.coli to co-culture with pathogenic Vibrio and create a prey-predator system, also called bio-control, to control Vibrio population in order to create an equilibrium between Vibrio and engineered E.coli. Therefore Vibrio population can be controlled in a particular concentration which will not able to infect marine organism such as shrimp, fish…etc."),a.default.createElement("div",null,a.default.createElement("div",{className:"illustration"},a.default.createElement("div",{className:"illust-image-wrapper"},a.default.createElement("img",{className:"illust-image",src:o.default.image.design.exp})),a.default.createElement("div",{className:"undertext"},"Fig 1. exponential growth of vibrio without constraint in fish farm")),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.design.equlibrium}),a.default.createElement("div",{className:"undertext"},"Fig 2. Vibrio population controlled by engineered E.coli forming a equilibrium")),a.default.createElement("div",null)),a.default.createElement("div",{className:"illustration"},a.default.createElement("div",{className:"illust-image-wrapper"},a.default.createElement("img",{className:"illust-image",src:o.default.image.design.shrimp})),a.default.createElement("div",{className:"undertext"},"Fig 3. Infected shrimp")),a.default.createElement("div",null,"Hence the goal of wetlab this year is going to design a gene circuit which can make engineered E.coli to secrete anti-vibrio substances[1] while vibrio concentration exceed hazard concentration, and engineered E.coli suicide[2] while vibrio concentration is low to avoid bio-safety concern."),a.default.createElement("div",{className:"illustration"},a.default.createElement("div",{style:{margin:"auto",width:"500px"}},a.default.createElement("img",{className:"illust-image",src:o.default.image.design.cycle}))),a.default.createElement("div",{className:"part"}," Our gene design can be divided into 3 parts. First is the “detection” part makes our E.coli sense the population of vibrio and respond to it.  Second, the “kill switch” part compares the level of 2 bacteria and switches our E.coli  between modes of secreting anti-vibrio substances and suicide substance. “Growth inhibitor” part includes a toxic protein (so called Colicin M) and a toxic small RNA (tsRNA)."),a.default.createElement("div",{className:"part"}," Though we want to conduct Vibrio related experiments in our lab, our lab belongs to BSL1 level and we can’t cultivate Vibrio. Therefore, we use 2 different kinds of E. coli. to simulate our succession model with one being as fake Vibrio, and another being as engineered E. coli. ")),c=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},"ACE is an abbreviation of “Automatic Color-detecting Examiner” which is able to provide a stable environment for any color-changing-related test. All of the processes are executed and the information of each color will be digitized automatically. Finally, we create our own database to store the data and analyze them more precisely. Combining with IoT technology, all of our data will be recorded and analyzed in our cloud database and be returned to our clients, which allows them to take control of the results."),a.default.createElement("div",{className:"part"},"In order to prove our ACE can be used in reality to solve the Vibrio problem in our society, we make a collaboration with the “Fisheries Research Institute” of the Taiwan government. 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OD 600 Reference point"),a.default.createElement("div",null,"LUDOX CL-X (45% colloidal silica suspension) is used as a single point reference to obtain a conversion factor to transform absorbance (Abs600) data from our plate reader into a comparable OD600 measurement as would be obtained in a spectrophotometer. We will get the data for OD600 of the H2O and LUDOX. The corrected Abs600 is calculated by subtracting the H2O reading. To convert measured Abs600 to OD600 is to let Reference OD600 divided by Abs600.")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"2. Particle Standard Curve"),a.default.createElement("div",null,"We prepared a dilution series of monodisperse silica microspheres and measured the Abs600 in a plate reader. The size and optical characteristics of these microspheres are similar to cells, and there was a known amount of particles per volume. This measurement allows us to construct a standard curve of particle concentration which can be used to convert Abs600 measurements to an estimated number of cells.")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"3. Fluorescence standard curve"),a.default.createElement("div",null,"Absolute fluorescence values cannot be directly compared from one instrument to another. In order to compare the fluorescence output of test devices between teams, it was necessary to create a standard fluorescence curve.")))),f=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},a.default.createElement("div",null,"First, we transformed Escherichia coli DH5α with these following plasmids."),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",null,"Negative control  BBa_R0040    Kit Plate 7 Well 2D"),a.default.createElement("div",null,"Positive control  BBa_I20270  Kit Plate 7 Well 2B"),a.default.createElement("div",null,"Test Device 1  BBa_J364000  Kit Plate 7 Well 2F"),a.default.createElement("div",null,"Test Device 2  BBa_J364001  Kit Plate 7 Well 2H"),a.default.createElement("div",null,"Test Device 3  BBa_J364002  Kit Plate 7 Well 2J"),a.default.createElement("div",null,"Test Device 4  BBa_J364007  Kit Plate 7 Well 2L"),a.default.createElement("div",null,"Test Device 5      BBa_J364008  Kit Plate 7 Well 2N"),a.default.createElement("div",null,"Test Device 6      BBa_J364009  Kit Plate 7 Well 2P"))),a.default.createElement("div",{className:"part"},"Secondly, two colonies from each plate were picked and inoculated in LB medium containing chloramphenicol overnight for 16-18 hours at 37°C and 220 rpm."),a.default.createElement("div",{className:"part"},"Finally, we measured the OD600 and fluorescence of transformed cells according to the protocol after 0, 6 hours. 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Outside of iGEM, he is now joining in an immunology laboratory and doing some researches about immune checkpoints, PD-1. Apart from the university studies, he is also active in diverse activities. For example, he has participated in several Chinese calligraphy contests in Taiwan and won many prizes.",contact:{facebook:"https://www.facebook.com/tysh0106",linkedin:"https://www.linkedin.com/in/tysh0106/"}},{name:"Kevin Wang",image:""+C.default.image.member.Kevin_Wang,position:"Wetlab",department:"Materials Science and Engineering/ Physics",description:"I am Kevin Wang, a sophomore student study in Material Sciences and Physics. \n      During iGEM, I design the gene circuit and most of gene fragments of our project this year, and including trouble shooting. This summer, I spent lots of time doing digestion-ligation, transform and PCR. Although failure make me feel frustration, I never give up till success. 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Also, Opera and EDM are the two music genres she loves most : )",contact:{facebook:"https://www.facebook.com/tinawu2406",linkedin:"https://www.linkedin.com/in/xin-fang-wu/",email:"tinawu2406@gmail.com"}},{name:"Ruby Chan",image:""+C.default.image.member.Ruby_Chan,position:"Wetlab",department:"Chemistry",description:"A girl who is never satisfied with the status quo.",contact:{facebook:"https://www.facebook.com/profile.php?id=100004026627188"}},{name:"Andy Yu",image:""+C.default.image.member.Kai_Chiang_Yu,position:"Wetlab",department:"Chemistry",description:"Andy, a senior student from department of chemistry, now struggling along the tenebrous road from hell.",contact:{facebook:"https://www.facebook.com/profile.php?id=100002090493215"}},{name:"Ricky Chan",image:""+C.default.image.member.Ricky_Chan,position:"Wetlab",department:"Applied Science",description:"A boy who is curious about this unknown world, and likes to connect with different people. 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Combining with IoT technology, all of our data will be recorded and analyzed in our cloud database and be returned to our clients, which allows them to take control of the results."),o.default.createElement("div",{className:"illustration"},o.default.createElement("div",{className:"illust-image-wrapper"},o.default.createElement("img",{className:"illust-image",src:s.default.image.drylab_overview.box}),o.default.createElement("div",{className:"undertext"},"Automatic Color-detecting Examiner")))),h=o.default.createElement("div",null,o.default.createElement("div",{className:"part"},o.default.createElement("div",null,"In order to prove our ACE can be used in reality to solve the problems in our society, we make a collaboration with the “Fisheries Research Institute” of the Taiwan government. They kindly provided us with their “Multiple-Vibrio Detecting Kit, MV-kit” and we integrated their MV-kit with our system to develop a brand-new",o.default.createElement("a",{href:"https://2018.igem.org/Team:NTHU_Taiwan/Application"}," Automatic Vibrio-detecting System"),".")),o.default.createElement("div",{className:"part"},"Through the optimization of ACE, we accelerate the detection rate of MV-kit. With our ACE, it becomes 2 times faster than the original MV-kit and the minimal detectable Vibrio concentration is about 100 CFU/ mL within 24 hours. In summary, we combine the traditional detecting methods with the cutting-edge technology to achieve a better effect. In addition, following with our IoT system makes the water quality testing system in aquatic agriculture industry step forward to the whole new world of “intelligent aquaculture”."),o.default.createElement("div",{className:"illustration"},o.default.createElement("img",{className:"illust-image",src:s.default.image.drylab_overview.plate}),o.default.createElement("div",{className:"undertext"},"The color gradient changing process"))),m=o.default.createElement("div",null,o.default.createElement("div",null,"Components list and cost on ",o.default.createElement("a",{href:" https://docs.google.com/spreadsheets/d/1NsoRao-FtywW1KCeFYrxMIjJG3kVGQRcg_fIhxjn7ZQ/edit?usp=sharing"},"Google drive")),o.default.createElement("div",null,"3D printing files on ",o.default.createElement("a",{href:"https://drive.google.com/drive/folders/1tPQEVtNHB_hzhalPlwZu4bzyQVSDUPkN?usp=sharing"},"Google Drive"))),g={viewbar:{Title:"Drylab",Subtitle:"ACE- Automatic Color-detecting Examiner",image:s.default.image.topfull.drylab_overview,Underimage:f},content:{Introduction:d,parts:[{Title:"ACE",Description:p},{Title:"Application",Description:h},{Title:"File download",Description:m}]}},v=(0,l.inject)("appState")(r=(0,l.observer)(r=function(e){function t(e){return function(e,t){if(!(e instanceof t))throw new TypeError("Cannot call a class as a function")}(this,t),function(e,t){if(!e)throw new ReferenceError("this hasn't been initialised - super() hasn't been called");return!t||"object"!=typeof t&&"function"!=typeof t?e:t}(this,(t.__proto__||Object.getPrototypeOf(t)).call(this,e))}return function(e,t){if("function"!=typeof t&&null!==t)throw new TypeError("Super expression must either be null or a function, not "+typeof t);e.prototype=Object.create(t&&t.prototype,{constructor:{value:e,enumerable:!1,writable:!0,configurable:!0}}),t&&(Object.setPrototypeOf?Object.setPrototypeOf(e,t):e.__proto__=t)}(t,a.Component),i(t,[{key:"render",value:function(){return o.default.createElement(u.default,g)}}]),t}())||r)||r;t.default=v},function(e,t,n){"use strict";Object.defineProperty(t,"__esModule",{value:!0});var r=function(){function e(e,t){for(var n=0;n<t.length;n++){var r=t[n];r.enumerable=r.enumerable||!1,r.configurable=!0,"value"in r&&(r.writable=!0),Object.defineProperty(e,r.key,r)}}return function(t,n,r){return n&&e(t.prototype,n),r&&e(t,r),t}}(),i=n(0),a=s(i),o=(s(n(10)),s(n(4))),l=s(n(8));function s(e){return e&&e.__esModule?e:{default:e}}var u=a.default.createElement("div",null,a.default.createElement("div",null,"The reaction platform mainly consists of three parts: "),a.default.createElement("div",{className:"innerpart"},"1. The 96-Well Microtitration Plate for sample storage and reaction "),a.default.createElement("div",{className:"innerpart"},"2. Light blocking base to black out the area we’re not using to make it easier for camera to detect the color changes of each well"),a.default.createElement("div",{className:"innerpart"},"3. Auto moving lid to prevent the reactive reagent from evaporating")),c=a.default.createElement("div",null,a.default.createElement("div",null,"After extracting a sample from the fish farm, the sample will be dripped into the 96 -well plate. In the plate, we put in the mixture of MV-kit buffer for the sample to react. There are several reasons we chose to use 96-well plates instead of eppendorf or 6/24/48 well plate."),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"innerpart"},"1. Eppendorf has an unremovable lid, which is not convenient for our auto sampling design."),a.default.createElement("div",{className:"innerpart"},"2. The base of eppendorf is sharp and liquid will not distribute evenly, which is not suitable for color sensing while 96-well plates are flat."),a.default.createElement("div",{className:"innerpart"},"3. The volume of the 96-well plate is small enough for us to put the sample in, while the other plates are too big."),a.default.createElement("div",{className:"innerpart"},"4. It can contain the most samples in one single plate. In that way, we can reduce the rate of changing new plates.")),a.default.createElement("div",{className:"illustration"},a.default.createElement("table",{className:"table"},a.default.createElement("tbody",null,a.default.createElement("tr",null,a.default.createElement("th",{className:"tablehead"},"Number of well"),a.default.createElement("th",{className:"tablehead"},"Radius (mm) "),a.default.createElement("th",{className:"tablehead"},"Max Volume (mL) "),a.default.createElement("th",{className:"tablehead"},"Growing Area (cm2)")),a.default.createElement("tr",null,a.default.createElement("td",{className:"tabledata"},"96"),a.default.createElement("td",{className:"tabledata"},"4.5 "),a.default.createElement("td",{className:"tabledata"},"0.36"),a.default.createElement("td",{className:"tabledata"},"0.16")),a.default.createElement("tr",null,a.default.createElement("td",{className:"tabledata"},"48"),a.default.createElement("td",{className:"tabledata"},"10.2"),a.default.createElement("td",{className:"tabledata"},"1.7"),a.default.createElement("td",{className:"tabledata"},"0.8")),a.default.createElement("tr",null,a.default.createElement("td",{className:"tabledata"},"24"),a.default.createElement("td",{className:"tabledata"},"15.6"),a.default.createElement("td",{className:"tabledata"},"3.4"),a.default.createElement("td",{className:"tabledata"},"1.9")),a.default.createElement("tr",null,a.default.createElement("td",{className:"tabledata"},"12"),a.default.createElement("td",{className:"tabledata"},"22.1"),a.default.createElement("td",{className:"tabledata"},"6.9"),a.default.createElement("td",{className:"tabledata"},"3.8")),a.default.createElement("tr",null,a.default.createElement("td",{className:"tabledata"},"6"),a.default.createElement("td",{className:"tabledata"},"34.8"),a.default.createElement("td",{className:"tabledata"},"16.8"),a.default.createElement("td",{className:"tabledata"},"9.5")))),a.default.createElement("div",{className:"undertext"},"reference: http://www.dxy.cn/bbs/topic/9584997"),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.platform.transparent})),a.default.createElement("div",{className:"undertext"},"reference: http://www.carelifetech.com.tw/?96%E5%AD%94%E7%9B%A4-96-well-plate,127"))),f=a.default.createElement("div",null,a.default.createElement("div",null,"Since we need to use the camera to detect the color changes of each well, we need to create an environment that can make the color as clear as possible.  Therefore, we use 3D printing and polylactic acid (PLA) to print out a platform that blacks out all the area except for the 24 holes of each plate that have our sample in."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.platform.block}))),d=a.default.createElement("div",null,a.default.createElement("div",null,"Since the MV-kit buffer and solution might evaporate, we need to construct an auto moving lid. The lid has 48 holes on it, each hole is 9.1 mm next to the well. This lid is connected with a gear and a stepper motor. Every time the sample is being dripped into the well, the motor will move the lid by 9.1 mm. The gear system and the base of the motor are also made by 3D printing. Moreover, we’ve designed the lid to be detachable so that the farmer can remove the lid when they are going to substitute the 96 well plates with a new one every two weeks. "),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.platform.moving}))),p={viewbar:{Title:"Platform",Subtitle:"Plate and the moving lid",image:o.default.image.topfull.platform},content:{Introduction:u,parts:[{Title:"96-Well Microtitration",Description:c},{Title:"Light blocking",Description:f},{Title:"Moving lid",Description:d}]}},h=function(e){function t(){return function(e,t){if(!(e instanceof t))throw new TypeError("Cannot call a class as a function")}(this,t),function(e,t){if(!e)throw new ReferenceError("this hasn't been initialised - super() hasn't been called");return!t||"object"!=typeof t&&"function"!=typeof t?e:t}(this,(t.__proto__||Object.getPrototypeOf(t)).apply(this,arguments))}return function(e,t){if("function"!=typeof t&&null!==t)throw new TypeError("Super expression must either be null or a function, not "+typeof t);e.prototype=Object.create(t&&t.prototype,{constructor:{value:e,enumerable:!1,writable:!0,configurable:!0}}),t&&(Object.setPrototypeOf?Object.setPrototypeOf(e,t):e.__proto__=t)}(t,i.Component),r(t,[{key:"render",value:function(){return a.default.createElement(l.default,p)}}]),t}();t.default=h},function(e,t,n){"use strict";Object.defineProperty(t,"__esModule",{value:!0});var r=function(){function e(e,t){for(var n=0;n<t.length;n++){var r=t[n];r.enumerable=r.enumerable||!1,r.configurable=!0,"value"in r&&(r.writable=!0),Object.defineProperty(e,r.key,r)}}return function(t,n,r){return n&&e(t.prototype,n),r&&e(t,r),t}}(),i=n(0),a=s(i),o=(s(n(10)),s(n(8))),l=s(n(4));function s(e){return e&&e.__esModule?e:{default:e}}var u=a.default.createElement("div",null,"For the results of color-changing, we write a program to detect them. In our device, there are 24 samples processing the chemical reaction simultaneously. During the reaction, we want to get all samples’ color values immediately, so we adopt image detection executed by Raspberry Pi to analyze the whole status directly. The following are three parts for our image detection, which is how to do, advantages, and problems respectively."),c=a.default.createElement("div",null,a.default.createElement("div",null,"The following is the materials image detection needs"),a.default.createElement("table",{className:"table"},a.default.createElement("tbody",null,a.default.createElement("tr",null,a.default.createElement("td",{className:"tabledata"},"Raspberry Pi 3 Model B v1.3"),a.default.createElement("td",{className:"tabledata"},"1")),a.default.createElement("tr",null,a.default.createElement("td",{className:"tabledata"},"Webcam Logitech C270"),a.default.createElement("td",{className:"tabledata"},"1")),a.default.createElement("tr",null,a.default.createElement("td",{className:"tabledata"},"Wide-angle lens 37mm 0.6X"),a.default.createElement("td",{className:"tabledata"},"1")))),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:l.default.image.imaging.materials})),a.default.createElement("div",{className:"part"},"The core of image detection is executed by Raspberry Pi and the program language we use is python. With a camera and wide-angle lens, it can capture the scene in a relatively lower height. To do the image processing, OpenCV, open source library, is the best choice. Because the program will send warning message to the users, for simplicity and stability, we choose MQTT to be the transport protocol."),a.default.createElement("div",{className:"part"},"The following three steps are the processes our image detection works on. First, it captures the scene using a camera in a specified time. Second, the image is analyzed immediately after one is captured and the results are record. Third, after the analysis, if there is a color-changing sample, it will send the warning message to the users. All the steps are operated recursively until all the samples are analyzed.")),f=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},"For our image detection, we have three advantages, which are accuracy, automation, and flexibility, respectively. With these three, it can match our goal of designing ACE, i.e., automation and also increase usability compared to other devices used in the laboratory nowadays."),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"1. Accuracy"),a.default.createElement("div",{className:"innerpart"},"Because our 24 samples are close to each other, the accuracy is necessary to avoid the wrong result of analysis. With image detection, we can set the section where we want to analyze corresponding to the samples on the picture. Therefore, if all samples are located in advance carefully, we can get high accuracy for the results.")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"2. Automation"),a.default.createElement("div",{className:"innerpart"},"Automation is our goal of designing the device. To achieve this, the program can automatically do the following things, i.e., capturing the scene, analyzing the captured image, recording the results, and sending the warning messages to users. With the four steps as mentioned above operated cyclically, the program achieves automation.")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"3. Flexibility"),a.default.createElement("div",{className:"innerpart"},"The section program analyzes on the captured image can be modified whenever the captured one is changed. Therefore, we don’t have to fix the position of all samples. Due to this advantage, any type of sample can be analyzed accurately."))),d=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},"During the development of image detection, we had made some choices to solve many problems, including the specific practice of detection, development board, the algorithm to analyze the images, and the environment light. The following describes each problem, and the reasons why we made these choices."),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"1. Image detection vs. Color sensor"),a.default.createElement("div",{className:"innerpart"},"Initially, color sensor is our direction to design the device. Moving the color sensor to the top of each sample can get the color of that. However, our samples are close to each other, and the resolution of a color sensor is not high enough to detect the color of single sample accurately, so the direction is changed to image detection with the camera capturing the scene. In this way, it analyzes the image directly, the accuracy increases by a large margin.")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"2. Raspberry Pi vs. Other development boards"),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:l.default.image.imaging.board})),"For the development boards, there are three choices, Arduino, Udoo Neo, and Raspberry Pi. Considering the efficiency, Arduino might not be able to run OpenCV effectively. The another consideration is the cost. Because we want to design a low-cost device cheaper than the similar machine sold on the market, the price of Udoo Neo is too expensive for us. Therefore, we chose Raspberry Pi to be our execution core.")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"3. Simplified location vs. Houghcircles"),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:l.default.image.imaging.algorithm})),"In the beginning, we planned to use OpenCV built-in function Houghcircles to do image detection. The detail of the function is that it can find circles on the image automatically. It completely meets our expectation. However, the distance and angle of each sample from the camera is different. As a result, the shape of the sample is not a circle on the captured image and leads to wrong detection by Houghcircles. For this reason, we changed the method and decided to locate the samples manually to get a high accuracy.")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"4. LED strip vs. Big bulb"),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:l.default.image.imaging.light})),"In order to prevent the influence of environment outside the device, ACE is designed to be enclosed. Therefore, it is completely dark inside the device and needs a light source to do image detection. In the beginning, we thought a big bulb is simple and suitable. Nevertheless, after we tested, we found the light on the image is not even, it would result in the wrong analysis. As a result, we chose LED strip. Its light is not only even but also bright enough.")),p=a.default.createElement("div",null,"You can download the source of our program on",a.default.createElement("a",{href:"https://github.com/igem-tw-nthu/opencv"}," Github")),h={viewbar:{Title:"Imaging",Subtitle:"Imaging by Raspberry Pi with OpenCV",image:l.default.image.topfull.imaging},content:{Introduction:u,parts:[{Title:"How to Do",Description:c},{Title:"Advantages",Description:f},{Title:"Problems",Description:d},{Title:"Code Download",Description:p}]}},m=function(e){function t(){return function(e,t){if(!(e instanceof t))throw new TypeError("Cannot call a class as a function")}(this,t),function(e,t){if(!e)throw new ReferenceError("this hasn't been initialised - super() hasn't been called");return!t||"object"!=typeof t&&"function"!=typeof t?e:t}(this,(t.__proto__||Object.getPrototypeOf(t)).apply(this,arguments))}return function(e,t){if("function"!=typeof t&&null!==t)throw new TypeError("Super expression must either be null or a function, not "+typeof t);e.prototype=Object.create(t&&t.prototype,{constructor:{value:e,enumerable:!1,writable:!0,configurable:!0}}),t&&(Object.setPrototypeOf?Object.setPrototypeOf(e,t):e.__proto__=t)}(t,i.Component),r(t,[{key:"render",value:function(){return a.default.createElement(o.default,h)}}]),t}();t.default=m},function(e,t,n){"use strict";Object.defineProperty(t,"__esModule",{value:!0});var r=function(){function e(e,t){for(var n=0;n<t.length;n++){var r=t[n];r.enumerable=r.enumerable||!1,r.configurable=!0,"value"in r&&(r.writable=!0),Object.defineProperty(e,r.key,r)}}return function(t,n,r){return n&&e(t.prototype,n),r&&e(t,r),t}}(),i=n(0),a=u(i),o=(u(n(10)),u(n(4))),l=u(n(8)),s=u(n(136));function u(e){return e&&e.__esModule?e:{default:e}}var c=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},a.default.createElement("div",null,"For most of the detection reagent, they can only react with the small amount of sample each time. Taking MV-kit (Multiple Vibrio Detecting Kit) that we’re using for vibrio detection for example, it needs to have fix amount of 35μl sample fluid being collected every time so that we can get the better result of the later on reaction with the reagent. We choose mini peristaltic pump to fulfill our need. There are a few advantages of choosing peristaltic pump as our dispensing media[1]:"),a.default.createElement("div",null,a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",{className:"part-title"},"1. Accurate metering "),a.default.createElement("div",null,"Peristaltic pumps use flexible tubing to run through rollers in the pump head. As fluid goes through the pump head, packet of fluids can be formed. Simply by determining the size of the tubing and the speed of the roller turning in the pump head, the accurate metering of the media can be achieved.")),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",{className:"part-title"},"2. Self priming "),a.default.createElement("div",null,"Peristaltic pumps can draw fluid into the tubing when starting dry while other pumps require user to fill pump and suction line with fluid before use which can be inconvenient and also create potential for contamination.")),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",{className:"part-title"},"3. Non-siphoning "),a.default.createElement("div",null,"A peristaltic pump is non-siphoning which means that it prevents back flow into the system when the pump stop turning. This promotes accuracy during dispensing.")))),a.default.createElement("div",{className:"part"},a.default.createElement("div",null,"As a result, peristaltic pumps are often used for applications involving metering, dispensing and precise general transfer. Here, we use it in our device and we get the stl. files from the open source of the YouTube channels called “electronic crafts”[2]. The dimension of our peristaltic pump is 31 x 31 x 11 (mm), and the detail of other components are showed in Table1 and Figure 1. We printed the main components out by using the SLA 3D printer."),a.default.createElement("table",{className:"table"},a.default.createElement("tbody",null,a.default.createElement("tr",null,a.default.createElement("th",{colSpan:"6",className:"tablehead"},"Peristaltic Pump")),a.default.createElement("tr",null,a.default.createElement("td",{className:"tabledata"},"Dimension"),a.default.createElement("td",{className:"tabledata"},"Tube diameter"),a.default.createElement("td",{className:"tabledata"},"Tube material"),a.default.createElement("td",{className:"tabledata"},"Number of rollers"),a.default.createElement("td",{className:"tabledata",style:{margin:"0px 10px"}},"Bearing"),a.default.createElement("td",{className:"tabledata"},"Motor")),a.default.createElement("tr",null,a.default.createElement("td",{className:"tabledata"},"31 x 31 x 11 (mm)"),a.default.createElement("td",{className:"tabledata"},a.default.createElement("div",null,"OD: 3mm"),a.default.createElement("div",null,"ID: 1.5mm")),a.default.createElement("td",{className:"tabledata"},"Silicone rubber"),a.default.createElement("td",{className:"tabledata"},"three"),a.default.createElement("td",{className:"tabledata"},a.default.createElement("div",null,"OD: 9mm"),a.default.createElement("div",null,"ID: 3mm")),a.default.createElement("td",{className:"tabledata"},a.default.createElement("div",null,"28BYJ-48"),a.default.createElement("div",null,"12V"))))),a.default.createElement("div",{className:"undertext"},"Table 1"),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.sampling.pump}),a.default.createElement("div",{className:"undertext"},"Figure 1"))),a.default.createElement("div",{className:"part"},"Because peristaltic pumps deliver fixed amounts of fluid with each pass of a roller over the tube, the size of the tube has a direct effect on the amount of fluid delivered. Well-designed pumps are engineered to work with an optimum tubing size or range of tubing sizes, which take into account both the tube's inside diameter and wall thickness. The inside diameter determines the amount of fluid delivered with each turn of the rotor, while the wall thickness affects the tubing's ability to spring back to its original shape after each compression, which has a great influence on the overall life of the tubing[3]."),a.default.createElement("div",{className:"part"},"We choose the tube with inner diameter 2mm and wall thickness 1mm. Three bearings are used as the rollers in our peristaltic pump. The outer diameter of the bearing is 9mm and the inside diameter is 3mm. The peristatic pump after assembling can be seen in Figure 2. We can also calculated the theoretical discharge to know the theoretical performance of the peristaltic pump by using the equation below[4],"),a.default.createElement("div",{className:"part",style:{display:"flex",alignItems:"center"}},a.default.createElement("div",null,a.default.createElement("div",null,a.default.createElement(s.default.Provider,null,a.default.createElement(s.default.Node,{inline:!0,formula:"flowrate (μl/s) = V \\cdot L \\cdot N \\cdot RPM \\cdot10^3 \\cdot 60"}))),a.default.createElement("div",null,"V= Volume of occluded tubing (mm^3/mm)"),a.default.createElement("div",null,"L= Tubing length that will be occluded by pump rollers (mm)"),a.default.createElement("div",null,"N= Number of rollers on the rotor."),a.default.createElement("div",null,"RPM= pump rpm")),a.default.createElement("div",{style:{marginLeft:"auto",padding:"0 20px"}},a.default.createElement("img",{style:{height:"180px"},src:o.default.image.sampling.full_pump}),a.default.createElement("div",{className:"undertext"},"Figure 2"))),a.default.createElement("div",{className:"part"},a.default.createElement("div",null,"We have done multiple testing and calculated the weight of each droplet by measuring the weight of every ten droplets to get the average value. The average weight of each droplet is 0.035+-0.00072μl, and the rest of the measuring records are showed in Figure 3. This is exactly what we need for sample preparing for the MV-kit in our experiments."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.sampling.list}),a.default.createElement("div",{className:"undertext"},"Figure 3"))),a.default.createElement("div",{className:"part",style:{display:"flex",alignItems:"center"}},a.default.createElement("div",{style:{marginRight:"auto",padding:"0px 30px"}},a.default.createElement("img",{style:{height:"180px"},src:o.default.image.sampling.head}),a.default.createElement("div",{className:"undertext"},"Figure 4")),a.default.createElement("div",null,"Although we can almost get the precise amount of sample each time, it’s still hard to control the later droplets from dropping down cause of the gravity. Hence, we combine the pipette tip (Figure 4) at the end of the silicon tube to reduce the diameter and slow down the flowing rate. In addition, we also design the special lid with slopes in between to cover on the 96-well plate. We can utilize the slope design not only to drop the first few droplets that might be contaminated by the previous sample but also use them as the tunnels to collect the waste sample and remove all of them after the whole examination is done.")),a.default.createElement("div",{className:"illustration"},a.default.createElement("video",{className:"illust-image",autoPlay:!0,muted:!0,loop:!0},a.default.createElement("source",{src:o.default.video.lid,type:"video/mp4"})))),f=a.default.createElement("div",null,a.default.createElement("div",null,"We use two 12V 28BYJ-48 stepper motors to control the XY axis to move our sample to the specific coordinate and connect them to Arduino as well as uln2003 motor drivers to control the speed and time. The step angle of the motor is 5.625 degrees and the reduction ratio is 1/64, therefore, for one revolution it needs 4096 steps. The diameter of the gear mounted on the motor is 36mm, so for each step, it can linearly move forward or backward 1.76mm. The length between two wells on 96-well plate is about 18mm, thus, this motor can absolutely satisfy our need for our low-cost XY plane setting. "),a.default.createElement("div",{className:"illustration"},a.default.createElement("video",{className:"illust-image",autoPlay:!0,muted:!0,loop:!0},a.default.createElement("source",{src:o.default.video.sampling,type:"video/mp4"})))),d=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},"At first, we’re trying to set up the XY axis by using the DC motor to control it in considering of the low-cost budget. However, we found out that since the DC motor is categorized as analog motor, we can only drive them by sending power signals (two wire) to the motor to drive them. This would be tricky for us to do the sophisticated planned moves because we won’t know exactly where they may be located except trying to predict the moves over time within a relative margin of error(3-8%)[5]. Moreover, the friction between the gear rack and the motor make it even hard to control and need to build up more complex moves manually. "),a.default.createElement("div",{className:"part"},"On the other hand, stepper motors are digital motors, we can actually send position signals (four wire) to the motor to drive them. It’s possible for us to know exactly where they are at all times, and would be easier for us to do ‘repeat’ motion passes and have it return to a ‘home and start’ position precisely. For this reason even most sophisticated of a moves can be achieved. "),a.default.createElement("div",{className:"part"},"Due to all the reasons above, instead of making motion decision ‘on the fly’ at the rig, we then choose stepper motor as our driving motor.")),p=a.default.createElement("div",null,a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.sampling.result}))),h=a.default.createElement("div",null,a.default.createElement("div",{className:"reference"},"1. https://www.manufacturingchemist.com/news/article_page/Peristaltic_pumps__advantages_and_applications/74693"),a.default.createElement("div",{className:"reference"},"2. Peristaltic pump design: https://www.youtube.com/watch?v=J2KLq7485kM"),a.default.createElement("div",{className:"reference"},"3. https://archive-resources.coleparmer.com/TechInfo/HTMLFiles/PPTube_match.htm"),a.default.createElement("div",{className:"reference"},"4. Suhas R. Dhumal, Design and Development of Rotary Peristaltic Pump, International Journal of Science and Advanced Technology (ISSN 2221-8386)"),a.default.createElement("div",{className:"reference"},"5. https://support.dynamicperception.com/hc/en-us/articles/202986595-What-is-the-difference-between-DC-motors-and-Stepper-motors-")),m={viewbar:{Title:"Sampling",Subtitle:"Auto sampling with peristaltic pump ",image:o.default.image.topfull.sampling},content:{Introduction:"In our device, in order to let the following detected process work out properly, we first set up the automated sampling mechanism. 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However, we found that it is not bright enough, so we chose another kind of light."),a.default.createElement("div",{className:"part"},"After this, we choose the white light bar and connect it with Arduino. Since the voltage afforded by Arduino is 12V, it’s too bright to observe, so we apply the voltage-gain-module to get the appropriate voltage. Then we find out the white light may sometimes be difficult for us to tell apart whether the liquid changes color or not. After discussion, we change the white light to yellow light and the problem is solved. ")),c=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},'Our Vibrio detection device needs to be able to react in a stable 30 ° C environment. It is hoped that a method can be designed to heat up when the ambient temperature is high and cool down when the temperature is low. The cooling technologies used today are mainly the following: "high pressure gas expansion and cooling", "liquid evaporation and refrigeration", "gas vortex cooling", "semiconductor refrigeration" and "chemical cooling", and we choose "semiconductor cooling", which is a relatively easy to complete and within the capability range.'),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"First step"),a.default.createElement("div",{className:"innerpart"},"Common methods for cooling semiconductors include water cooling and fan cooling. Among them, due to our large architecture, water cooling is effective but too cumbersome and heavy. Limited by the space configuration, we use fan cooling. (Picture 4) By keep bringing the heat away, the temperature gradient of the chip become larger, and finally it will stop at a balanced point. We also add a fan on the cold side since we want the cold air to be uniformed inside the box. (Picture 5) Temperature-controlled circuits are more cumbersome and too complicated circuits are not suitable for use in our system, so we try to use temperature control module first. (Picture 6)"),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:l.default.image.environment.first}),a.default.createElement("div",{className:"undertext"})),a.default.createElement("div",{className:"innerpart"},"We found that as we put the cooling device in a cardboard box for testing, the result is not good since the space is not sealed and made the chip hard to cool down the environment."),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",{className:"innerpart"},"Needed materials:"),a.default.createElement("div",null,"two 5V-2wire fans, two small heat sinks, some thermal grease, one thermoelectric cooling chip provided by a national competition in Taiwan, temperature control module."))),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Second step"),a.default.createElement("div",{className:"innerpart"},"To enlarge the temperature difference, we add a more powerful fan on the hot side to make sure the rate of heat dissipation is enough for the rate of chip getting cold, which is a 12V-2wire one. As a result, the inbox environment could drop 3℃, but still didn’t reach our expectation."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:l.default.image.environment.second}),a.default.createElement("div",{className:"undertext"},a.default.createElement("div",null,"Figure 1: The air temperature inside the box drops from 26.6℃ to about 18℃ in 7 minutes, and maintains at 18℃ for 30 minutes. (cold device) The air temperature inside the box rises from 27.2℃ to 32.8℃ in about 6 minutes, and maintains at 32.8℃ for 15 minutes. (hot device)"),a.default.createElement("div",null,"Figure 2: When cold devices turn off, the air temperature inside the box rises from 27.2℃ to 32.8℃ in about 6 minutes, and maintains at 32.8℃ for 15 minutes. (cold device) When the hot device turn off, the air temperature inside the box drops from about 32.8℃ to 27.6℃ in 6 minutes, the same as the time to heat up the air. (hot device)"))),a.default.createElement("div",{className:"innerpart"},'During our experiment, we found that the temperature control module couldn’t provide a two-state system, which is hard to be designed in our project. Therefore, we try another way by using Arduino and relay module to control the temperature directly. Relay, also known as electric cymbal, is an electronic control device that has a control system (also known as an input circuit) and a controlled system (also known as an output circuit). It is usually used in automatic control circuits. It is actually used as an "automatic switch" that uses a small current to control large currents. Therefore, it plays the role of automatic adjustment, safety protection and conversion circuit in the circuit. Besides, we chose an IC called LM35 as our temperature sensor by transforming resistance into temperature. The wiring diagram is as below. (Picture 8)'),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",{className:"innerpart"},"Needed materials: "),"Cold device is combined by 12V-2wire fan, heat sink, thermal grease, thermoelectric cooling chip, thermal grease, heat sink, 5V-2wire fan (from up to down); Temperature controlling: Arduino for control, LM35 for sensing, relay module for switch.")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Final step"),a.default.createElement("div",{className:"innerpart"},"To reduce the unknown impact on the experiment, we use two cold device and one hot device to control the temperature. Hot device is similar to cold one, the only different is its two fans are all 5V-2wire, which is the device we used in the first period."),"  ",a.default.createElement("div",{className:"innerpart"},"This time we use two cold devices to find for the lowest temperature. Figure 1 shows the result we test cold or hot devices, and Figure 2 shows the changing after we turn off the devices."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:l.default.image.environment.final}),a.default.createElement("div",{className:"undertext"})),a.default.createElement("div",{className:"inner-part"},"After discussion, we decide to give styrofoam a try, we add it inside the box and do the rest of the experiment, includes add it or not add it. We finally decide to use two cold devices and one hot device without adding any material."),a.default.createElement("table",null),a.default.createElement("div",{className:"innerpart"},"We use Arduino connects with relay and temperature sensor to control the temperature. When the sensor detects the temperature lower than the setting, hot device is activated, it would heat up the air inside the device until it reaches the target temperature. Then, the hot device continues working that makes the temperature higher than the ideal one, this time, the cold devices turn on and cool down the environment. At the end, the temperature would be controlled in the range we have written in Arduino."),a.default.createElement("div",{className:"innerpart"},"In the beginning, we set the temperature to 27℃, which is a good condition for germs to grow. However, we neglect that there would be some the delay of temperature change between the air and the sensor, and the temperature around the sensor became higher than the environment, which affected the detecting correction. After we tidied up the wires, the problems disappeared and came into perfect."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:l.default.image.environment.end}),a.default.createElement("div",{className:"undertext"},"Figure 6: Temperature control simulation. The hot device turns on at the beginning, and the cold devices turn on when the temperature reaches 28.3℃, in about 6 minutes.")))),f=a.default.createElement("div",null,a.default.createElement("div",{className:"reference"},"1. Thermal Conductivity and Thermoelectric Power of Semiconductors. I. Terasaki, in Reference Module in Materials Science and Materials Engineering, 2016"),a.default.createElement("div",{className:"reference"},"2. Physics and Fundamental Theory. I. Terasaki, in Comprehensive Semiconductor Science and Technology, 2011"),a.default.createElement("div",{className:"reference"},"3. Thermoelectric cooling. JoséMeseguer, IsabelPérez-Grande, AngelSanz-Andrés, in Spacecraft Thermal Control, 2012"),a.default.createElement("div",{className:"reference"},"4. https://en.wikipedia.org/wiki/Thermoelectric_effect#/media/File:Thermoelectric_Cooler_Diagram.svg")),d={viewbar:{Title:"Environment",Subtitle:"Led Light, and peltier cooler",image:l.default.image.topfull.environment},content:{Introduction:"In order to make sure the observation results are stable and reliable, it’s necessary to control the experiment environment. Besides putting all the instruments inside our acrylic-made box (size: 20*30*35, unit: cm3), temperature and brightness are also important. In this case, we separate the “Environment” part into two sub-titles, temperature, and light. We will introduce more details in the following statements.",parts:[{Title:"Light",Description:u},{Title:"Temperature",Description:c},{Title:"Reference",Description:f}]}},p=function(e){function t(e){return function(e,t){if(!(e instanceof t))throw new TypeError("Cannot call a class as a function")}(this,t),function(e,t){if(!e)throw new ReferenceError("this hasn't been initialised - super() hasn't been called");return!t||"object"!=typeof t&&"function"!=typeof t?e:t}(this,(t.__proto__||Object.getPrototypeOf(t)).call(this,e))}return function(e,t){if("function"!=typeof t&&null!==t)throw new TypeError("Super expression must either be null or a function, not "+typeof t);e.prototype=Object.create(t&&t.prototype,{constructor:{value:e,enumerable:!1,writable:!0,configurable:!0}}),t&&(Object.setPrototypeOf?Object.setPrototypeOf(e,t):e.__proto__=t)}(t,i.Component),r(t,[{key:"render",value:function(){return a.default.createElement(o.default,d)}}]),t}();t.default=p},function(e,t,n){"use strict";Object.defineProperty(t,"__esModule",{value:!0});var r=function(){function e(e,t){for(var n=0;n<t.length;n++){var r=t[n];r.enumerable=r.enumerable||!1,r.configurable=!0,"value"in r&&(r.writable=!0),Object.defineProperty(e,r.key,r)}}return function(t,n,r){return n&&e(t.prototype,n),r&&e(t,r),t}}(),i=n(0),a=s(i),o=s(n(4)),l=s(n(8));function s(e){return e&&e.__esModule?e:{default:e}}var u=a.default.createElement("div",null,a.default.createElement("div",null,"In order to achieve real time data analysis , when ACE detects the color-changing sample, it will send a message to the user. Between ACE and user’s mobile device, it needs a transfer protocol to communicate with each other. Here, we choose MQTT (Message Queuing Telemetry Transport) to upload the vibrio concentration onto the cloud. It is a lightweight messaging protocol for small sensors and mobile devices, optimized for high-latency or unreliable networks.  Therefore, this kind of method is useful in remote area where network bandwidth is limited, especially for the fish farm in Taiwan. We’ve designed an app for farmers to obtain information in time. In our app, we separate into three parts: Pond’s condition, Suggestions, and Multiple Pond map. "),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.software.app}))),c=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},"We’ve divided different bacteria’s concentration into three levels, Safe, Warning, and Danger. The standard of this division is refer to the advice of the Fisheries Research Center of Republic of China. After visiting the Marine Life museum Research center, the researcher also tell us that usually the concentration for vibrio to be pathogenic is >104CFU/ml."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.software.line})),a.default.createElement("div",{className:"part"},"Furthermore, in order to let the farmers keep track of how the vibrio in their farm has been growing, we’ve made a history record chart, of which the Y coordinate is concentration, and the X coordinate is time. The concentration points will be drawn every hour.  The chart can be drag, zoom in and out for a closer look. By knowing the condition of their pond beforehand and even predict how the amount of the vibrios might grow, the farmers can do precaution in order to prevent vibrios from harming their fish or shrimp.")),f=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},"When the concentration of vibrios is under the safety standard, the fisheries research center recommend the farmers to change the water environment, or adjust the amount of feed. Only when the condition is in danger that they can add the medicine.  "),a.default.createElement("div",{className:"part"},"In addition, we will send notification bar through android phones to farmers if the amount of vibrios exceed standard."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.software.warn}))),d=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},"The reason why we want to combine google map into our APP is that one farmer usually own more than one pond.  Then if they want to keep track of different pond, they need to number their farm. Most of the time, it’s difficult for farmers to memorize which pond does this number really referring to, which indicate that the managing system is not so user friendly. "),a.default.createElement("div",{className:"part"},"In our map, farmers can locate its position and to help its pond create a name.  Next time when they want to inspect the condition of each pond, they can just press on the name label on the map. In order to make the system more intuitive, we add in the function of turning the normal map into 3D map and satellite(RealView) one, which is more close to the real view on the ground."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.software.map}))),p={viewbar:{Title:"Software",Subtitle:"An app for real-time analysis of pond’s condition",image:o.default.image.topfull.software},content:{Introduction:u,parts:[{Title:"Pond’s condition",Description:c},{Title:"Suggestions",Description:f},{Title:"Multiple pond management map",Description:d}]}},h=function(e){function t(){return function(e,t){if(!(e instanceof t))throw new TypeError("Cannot call a class as a function")}(this,t),function(e,t){if(!e)throw new ReferenceError("this hasn't been initialised - super() hasn't been called");return!t||"object"!=typeof t&&"function"!=typeof t?e:t}(this,(t.__proto__||Object.getPrototypeOf(t)).apply(this,arguments))}return function(e,t){if("function"!=typeof t&&null!==t)throw new TypeError("Super expression must either be null or a function, not "+typeof t);e.prototype=Object.create(t&&t.prototype,{constructor:{value:e,enumerable:!1,writable:!0,configurable:!0}}),t&&(Object.setPrototypeOf?Object.setPrototypeOf(e,t):e.__proto__=t)}(t,i.Component),r(t,[{key:"render",value:function(){return a.default.createElement(l.default,p)}}]),t}();t.default=h},function(e,t,n){"use strict";Object.defineProperty(t,"__esModule",{value:!0});var r=function(){function e(e,t){for(var n=0;n<t.length;n++){var r=t[n];r.enumerable=r.enumerable||!1,r.configurable=!0,"value"in r&&(r.writable=!0),Object.defineProperty(e,r.key,r)}}return function(t,n,r){return n&&e(t.prototype,n),r&&e(t,r),t}}(),i=n(0),a=s(i),o=s(n(8)),l=s(n(4));function s(e){return e&&e.__esModule?e:{default:e}}var u=a.default.createElement("div",null,a.default.createElement("div",null,"The importance of damage caused by Vibrio would not be elucidated here. You can see the detailed information in",a.default.createElement("a",{href:"https://2018.igem.org/Description"}," Description"),"."),a.default.createElement("div",{className:"part"},a.default.createElement("div",null,"Nowadays, there are three possible scenarios for aquatic agriculture to face pathogenic Vibrio:"),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",null,"1. The infection of Vibrio is usually happening all of a sudden. So when fisherman aware of the infection, it often causes significant damage."),a.default.createElement("div",null,"2. The conventional Vibrio testing method is time-consuming and manpower-consuming."),a.default.createElement("div",null,"3. The current water quality testing equipment is mainly focusing on the physical and chemical index, there are no devices for testing bacteria"))),a.default.createElement("div",{className:"part"},a.default.createElement("div",null,"However, our automatic Vibrio-detecting system is able to solve these problems."),a.default.createElement("div",{style:{display:"flex",alignItems:"flex-end"}},a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:l.default.image.application.tcbs}),a.default.createElement("div",{className:"undertext"},"TCBS medium")),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:l.default.image.application.plate}),a.default.createElement("div",{className:"undertext"},"MV-kit"))))),c=a.default.createElement("div",null,a.default.createElement("div",null,"MV-kit was developed by Fisheries Research Institute in Taiwan. It’s a kind of selective medium for Vibrio and it can detect 17 different aquatic pathogenic Vibrio. Through observing the time of color-changing, we can know the approximate Vibrio concentration in the sample. After our modification, within 24 hours, the minimal detectable Vibrio concentration is 100 CFU/ mL. Take Vibrio harveyi for example:"),a.default.createElement("div",{style:{display:"flex",alignItems:"flex-end"}},a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:l.default.image.application.concentration}),a.default.createElement("div",{className:"undertext"},"CFU vs. O.D.")),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:l.default.image.application.time}),a.default.createElement("div",{className:"undertext"},"CFU vs. Time of color-changing"))),a.default.createElement("div",null,"We build a relationship of O.D. value and the amount of bacteria via spreading method. Then, using O.D. value as a reference point, we build another relationship between the color-changing time of MV-kit and the amount of bacteria. Accordingly, we can get a significant correlation of bacteria concentration and the color-changing time of MV-kit")),f=a.default.createElement("div",null,a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:l.default.image.application.flow})),a.default.createElement("div",{className:"part"},"We provide a complete service for the prevention of Vibrio-infection. Our ACE collects the water sample from the fish pond and drips it into the plate automatically. Simultaneously, the device provides a closed environment with constant temperature for the reaction. Also, the device monitors the color-changing time at all times. As long as the color changes and achieves the condition we have set up, the data will be recorded and be uploaded to the server. After analyzing, the final result will be transferred to the clients’ cellphone applications. If the concentration of Vibrio overtakes the minimal hazardous value which is 1000 CFU/ mL, the application will warn the fisherman."),a.default.createElement("div",{className:"part"},"Through our system, our clients can take control of the situation of their fish ponds and use our engineered E. coli or other plans to solve the problems.")),d=a.default.createElement("div",null,a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:l.default.image.application.stp})),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"1. Segmentation"),a.default.createElement("div",null,a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",{className:"part-title"},"Merchandises in current market have to face some difficulties:"),a.default.createElement("div",null,"The water quality detecting systems in the current market are merely focusing on the particular physical and chemical indexes in the waters, for example, pH value, redox potential, dissolved oxygen, etc. They determine the water quality by those indexes, however, the change of situation in the water is quite fast and complex. We cannot receive the correct and real-time information of Vibrio in the water. When we aware of the abnormality, the pathogenic Vibrio usually causes the death of fish and shrimps significantly.")),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",{className:"part-title"},"Long-term detection with high specificity:"),a.default.createElement("div",null,"The MV-kit developed by Fisheries Research Institute is able to detect the Vibrio in the water sample quantitatively and with high specificity. Our system can automatically monitor the change of Vibrio concentration over a long period of time. Our clients are able to notice the condition in the early stage of Vibrio growth and adopt the solutions.")),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",{className:"part-title"},"A real-time and automatic system:"),a.default.createElement("div",null,"Our mechanism design allows the device to collect the sample automatically and continuously. Furthermore, we also program an application which analyzes the Vibrio concentration automatically. Last but not least, we achieve the effect of real-time monitoring and report through IoT technology.")),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",{className:"part-title"},"Fast, simple, and low-cost:"),a.default.createElement("div",null,"The mechanism of our reaction is the simple color-changing process. Compared to the conventional Vibrio culture methods, our clients haven’t to equipped with the professional skills of bacteria culture, also, the process of testing becomes safer and faster. In comparison with the water quality testing system in the market, our system is low-cost and is able to express the Vibrio concentration directly and accurately."))),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:l.default.image.application.vs}))),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"2. Targeting"),a.default.createElement("div",null,a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",null,"Aquatic agriculture industry:"),a.default.createElement("div",null,"Aquatic agriculture is an important industry and it stimulates the economic growth in Taiwan every year. However, the Vibrio-related infection leads to inestimable economic damage locally and globally. Our system aims to provide real-time, long-term, and low-cost monitoring toward Vibrio. Through the automatic system, our clients can reduce the manpower-cost and ensure their fish ponds are safe at any time. In addition, our mechanism design is quite simple so that the maintenance cost is also quite low. The only thing fisherman has to do is change the consumable materials regularly.")),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",{className:"part-title"},"Suppliers of the water quality testing system:"),a.default.createElement("div",null,"Currently, there are several water quality testing system vendors in Taiwan, but there is no any company provides the automatic Vibrio-detecting system. Our device is easy to be integrated with other related systems to reduce the vendors’ research and development cost. In this way, we believe that we can establish a better platform and to create a win-win situation.")))),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"3. Positioning"),a.default.createElement("div",null,a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",{className:"part-title"},"Intelligent fishery:"),a.default.createElement("div",null,a.default.createElement("div",null,"With the advance of science and technology, the IoT popularizes gradually in current society. Besides, the traditional agriculture industries transform into the so-called “agriculture 4.0”. "),a.default.createElement("div",null,"Let’s take an in-depth look at aquatic agriculture in Taiwan. Our country is famous for its aquatic agriculture industry, however, the conventional way is quite time-consuming. Additionally, following with the global warming, the aquatic agriculture faces a huge challenge. In the train of those facts, the industrial population structure becomes imbalance and thus, exerting a crucial impact on the future of industrial development ultimately."),a.default.createElement("div",null,"In order to turn the tables, the concept and development of intelligent fishery are emerging in recent years. It leads our traditional fishery industry into a new era, raising the operational efficacy, the income, and reducing the cost. Based on the concept, our system is expected to solve the Vibrio-monitoring problems and to establish a database to find an optimized solution."))),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",{className:"part-title"},"Future resolution:"),a.default.createElement("div",null,"Hopefully, we can create a complete detecting platform for different pathogenic bacteria in the future. With low-cost and high-efficiency, our platform is able to minimize the economic damage result from the bacterial infection."))))),p={viewbar:{Title:"Application",Subtitle:"Automatic Vibrio-detecting System",image:l.default.image.topfull.application},content:{Introduction:u,parts:[{Title:"MV-kit",Description:c},{Title:"Automatic Vibrio-detecting System",Description:f},{Title:"Commercial Analysis",Description:d}]}},h=function(e){function t(){return function(e,t){if(!(e instanceof t))throw new TypeError("Cannot call a class as a function")}(this,t),function(e,t){if(!e)throw new ReferenceError("this hasn't been initialised - super() hasn't been called");return!t||"object"!=typeof t&&"function"!=typeof t?e:t}(this,(t.__proto__||Object.getPrototypeOf(t)).apply(this,arguments))}return function(e,t){if("function"!=typeof t&&null!==t)throw new TypeError("Super expression must either be null or a function, not "+typeof t);e.prototype=Object.create(t&&t.prototype,{constructor:{value:e,enumerable:!1,writable:!0,configurable:!0}}),t&&(Object.setPrototypeOf?Object.setPrototypeOf(e,t):e.__proto__=t)}(t,i.Component),r(t,[{key:"render",value:function(){return a.default.createElement(o.default,p)}}]),t}();t.default=h},function(e,t,n){"use strict";Object.defineProperty(t,"__esModule",{value:!0});var r=function(){function e(e,t){for(var n=0;n<t.length;n++){var r=t[n];r.enumerable=r.enumerable||!1,r.configurable=!0,"value"in r&&(r.writable=!0),Object.defineProperty(e,r.key,r)}}return function(t,n,r){return n&&e(t.prototype,n),r&&e(t,r),t}}(),i=n(0),a=s(i),o=s(n(8)),l=s(n(4));function s(e){return e&&e.__esModule?e:{default:e}}var u=a.default.createElement("div",null,a.default.createElement("div",null,"In order to control Vibrio cell-population density and maintain a bio-friendly environment in a sustainable way, we introduce the concept of biocontrol, which is known as the prey-predator model, to our system. We create a system that can automatically detect, compare, and control the cell-population density. The system is composed of three parts, which are a detector, comparator, and controller respectively (Fig 1). These three parts are connected together in the same gene circuit and function sequentially so that the concentration of Vibrio in the surroundings can be kept in a specific range. Then, a succession model can be achieved."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:l.default.image.wetlab_overview.system}),a.default.createElement("div",{className:"undertext"},"Fig 1. Component of cell population controlling system."))),c=a.default.createElement("div",null,"Our gene circuit design can be divided into three parts described down below."),f=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},"The Detector module is responsible for detecting the concentration of Vibrio species in the surroundings. We found that most of pathogenic Vibrio use quorum sensing to communicate with other individuals. Lux is one of the Vibrio quorum sensing systems. With Lux system, the Vibrio has the ability to secrete and detect specific chemicals, acyl homoserine lactones (AHLs, Fig2). Signal transduction relies on AHLs, and bacteria respond to it with different gene expression."),a.default.createElement("div",{className:"part"},"We use Lux gene originally isolated from Vibrio fischeri as our AHL sensor. The gene system is composed of LuxI, LuxR, and pLux. LuxI is a synthase that converts S-Adenosyl methionine (SAM) into one of specific AHLs, 3OC6HSL (Fig2). LuxR is an activating protein, and it will stimulate the transcription from Lux promoter (pLux) when 3OC6HSL binds to it.[1] We modify LuxI with RFP reporter to the E. coli. playing the role of Vibrio. We also modify LuxR and pLux into our genetically engineered E. coli., the cell-population controlling system, so that AHL level in the environment can be detected and the Vibrio population can be known (Fig3).        "),a.default.createElement("div",{className:"illustration"},a.default.createElement("div",{className:"illust-image-wrapper"},a.default.createElement("img",{className:"illust-image",src:l.default.image.wetlab_overview.ahl})),a.default.createElement("div",{className:"undertext"},"Fig 2. Chemical structure of 3-oxo-C6-HSL.")),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:l.default.image.wetlab_overview.detection}),a.default.createElement("div",{className:"undertext"},"Fig 3. An AHL detection scheme showing that AHL-LuxR complex activates the gene downstream from pLux."))),d=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},"The comparator module is responsible for comparing the cell population density of Vibrio with our engineered E. coli. to see if it exceeds the pathogenic level. It receives information getting from the detector module. If the concentration of Vibrio is higher than engineered E. coli., which means LuxR-AHL complex is present, the anti-Vibrio gene will be transcribed. On the other hand, if the concentration of engineered E. coli. is higher than Vibrio, which means LuxR-AHL complex is absent, the suicide gene will be transcribed to kill E. coli. itself. "),a.default.createElement("div",{className:"part"},"Our initial effort is to design a gene circuit contain LacR to accomplish our goal. However, it turns out that there are some problems and drawbacks. While AHL presences, LacR should be produced and bound to pLac promoter to inhibit suicide gene being expressed. Yet, LacR can’t 100 % bind to pLac and this happens in most operons, so-called leakage problem. Suicide gene may be expressed, and E. coli. will suicide even at high AHL concentration. This would cause the system to be collapsed, and the result became out of our expectations (Fig 4)."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:l.default.image.wetlab_overview.lacl}),a.default.createElement("div",{className:"undertext"},"Fig 4. Our initial design using Lac operon to regulate genes expression.")),a.default.createElement("div",{className:"part"},"We come up with a better solution for our kill switch by using riboswitch and Small Transcriptional-Activating RNA (STAR) to regulate our gene. STAR is a small RNA with its sequence completely complementary to riboswitch. Riboswitch is an intrinsic transcription terminator, a DNA segment with special secondary structure. It forms a hairpin structure and inhibits transcriptions of the genes downstream.[2] When STAR binds to the riboswitch, it will be turned on (Fig 5, 6). Current studies show that riboswitch can regulate protein better than operon because of the higher affinity between STAR and riboswitch. In addition, STAR is smaller than regular proteins and it degrades faster than regular protein, therefore can respond to environmental change more rapidly than operon."),a.default.createElement("div",{className:"illustration"},a.default.createElement("div",{className:"illust-image-wrapper"},a.default.createElement("img",{className:"illust-image",src:l.default.image.wetlab_overview.riboswitch})),a.default.createElement("div",{className:"undertext"},"Fig 5. The secondary structure of riboswitch and STAR")),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:l.default.image.wetlab_overview.star}),a.default.createElement("div",{className:"undertext"},"Fig 6. STAR turns on the target and activates the downstream transcriptional process")),a.default.createElement("div",{className:"part"},"We also use a small RNA segment called anti-STAR in our design. Its sequence is totally complementary to STAR, but has higher affinity than riboswitch does. When anti-SATR exists, STAR will bind to it prior to riboswitch. Consequently, when Lux-AHL triggers the transcription of anti-STAR, it binds to STAR and the riboswitch is turned off (Fig 7). When Lux-AHL is absent and anti-STAR isn’t generated, STAR will turn on the riboswitch."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:l.default.image.wetlab_overview.anti}),a.default.createElement("div",{className:"undertext"},"Fig 7. Anti-STAR makes STAR lose it functionality. The gene downstream is still inhibited by the target."))),p=a.default.createElement("div",null,a.default.createElement("div",null,"We use two growth inhibitors to control populations of Vibrio and engineered E. coli. One is for inhibiting Vibrio, and another is for inhibiting engineered E. coli. At first, we would like to use anti-Vibrio substances in our design. However, our lab belongs to Bio-safety Level 1 (BSL1) and we can’t use real Vibrio in our experiments to test the toxicity of anti-Vibrio substance due to bio-safety concern. Therefore, we use colicin M to kill E. coli. (fake Vibrio) in order to prove our bio-control model. On the other hand, we use toxin small RNA (tsRNA) and endolysin as suicide substances to kill engineered E. coli. itself."),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"1. Colicin M"),a.default.createElement("div",null,"Colicin M blocks the biosynthesis of peptidoglycan (murein) and O-antigen. Colicin M contains activity (cma) and immune gene (cmi). The product of cmi is an immune protein, preventing colicin M to inhibit cell wall synthesis by binding to and thus neutralizing it. In addition, most colicins are released by autolysis, but colicin M lacks a lysis gene.[3-4] It has less tendency to cause the death of producer cells, so it’s a suitable protein for the purpose of killing other bacteria without harming producer cells.")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"2. Toxic small RNA (tsRNA)"),a.default.createElement("div",null,"Toxic small RNAs are small non-coding RNA. We use the tsRNA isolated from Burkholderia cenocepacia. The Current study showed that it is toxic to the E. coli. host and reproducibly inhibit E. coli growth. TsRNA harbored a sequence that was complementary to the consensus ribosomal binding site (RBS). It is hypothesized that it might target and misregulate essential genes within the E. coli cell. TsRNA cannot penetrate the bacterial cell from the outside, so it is unable to affect bacteria except E. coli. host cells.[5] Additionally, producing tsRNA doesn’t require a translational process, thus the response time is more rapid. As a result, tsRNA become an ideal suicide substance for our design.")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"3. Endolysin"),a.default.createElement("div",null,"Endolysin is a peptidoglycan hydrolase that is secreted by double-stranded DNA lambda phage to comprise the bacterial cell wall. It causes cell lysis by disrupting murein structure.")),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:l.default.image.wetlab_overview.mechanism}),a.default.createElement("div",{className:"undertext"},"Fig 8. The overall gene circuit design"))),h=a.default.createElement("div",null,a.default.createElement("div",{className:"innerpart"},"1. Visick, K. L.; Foster, J.; Doino, J.; McFall-Ngai, M.; Ruby, E. G., Vibrio fischeri lux Genes Play an Important Role in Colonization and Development of the Host Light Organ. Journal of Bacteriology 2000, 182 (16), 4578-4586."),a.default.createElement("div",{className:"innerpart"},"2. Chappell, J.; Takahashi, M. K.; Lucks, J. B., Creating small transcription activating RNAs. Nature chemical biology 2015, 11 (3), 214-20."),a.default.createElement("div",{className:"innerpart"},"3. Harkness, R. E.; Olschlager, T., The biology of colicin M. FEMS microbiology reviews 1991, 8 (1), 27-41."),a.default.createElement("div",{className:"innerpart"},"4. Zeth, K.; Römer, C.; Patzer, S. I.; Braun, V., Crystal Structure of Colicin M, a Novel Phosphatase Specifically Imported by Escherichia coli>. The Journal of Biological Chemistry 2008, 283 (37), 25324-25331."),a.default.createElement("div",{className:"innerpart"},"5. Kimelman, A.; Levy, A.; Sberro, H.; Kidron, S.; Leavitt, A.; Amitai, G.; Yoder-Himes, D. R.; Wurtzel, O.; Zhu, Y.; Rubin, E. M.; Sorek, R., A vast collection of microbial genes that are toxic to bacteria. Genome research 2012, 22 (4), 802-9.")),m={viewbar:{Title:"Wetlab",Subtitle:"Detector, comparator, and controller module",image:l.default.image.topfull.wetlab_overview},content:{Introduction:u,parts:[{Title:"Cell Population Controlling System",Description:c},{Title:"Detector",Description:f},{Title:"Comparator",Description:d},{Title:"Controller",Description:p},{Title:"Reference",Description:h}]}},g=function(e){function t(){return function(e,t){if(!(e instanceof t))throw new TypeError("Cannot call a class as a function")}(this,t),function(e,t){if(!e)throw new ReferenceError("this hasn't been initialised - super() hasn't been called");return!t||"object"!=typeof t&&"function"!=typeof t?e:t}(this,(t.__proto__||Object.getPrototypeOf(t)).apply(this,arguments))}return function(e,t){if("function"!=typeof t&&null!==t)throw new TypeError("Super expression must either be null or a function, not "+typeof t);e.prototype=Object.create(t&&t.prototype,{constructor:{value:e,enumerable:!1,writable:!0,configurable:!0}}),t&&(Object.setPrototypeOf?Object.setPrototypeOf(e,t):e.__proto__=t)}(t,i.Component),r(t,[{key:"render",value:function(){return a.default.createElement(o.default,m)}}]),t}();t.default=g},function(e,t,n){"use strict";Object.defineProperty(t,"__esModule",{value:!0});var r=function(){function e(e,t){for(var n=0;n<t.length;n++){var r=t[n];r.enumerable=r.enumerable||!1,r.configurable=!0,"value"in r&&(r.writable=!0),Object.defineProperty(e,r.key,r)}}return function(t,n,r){return n&&e(t.prototype,n),r&&e(t,r),t}}(),i=n(0),a=u(i),o=u(n(4)),l=u(n(8)),s=u(n(136));function u(e){return e&&e.__esModule?e:{default:e}}var c=a.default.createElement("div",null,a.default.createElement("div",{className:"illustration"},a.default.createElement("div",{style:{margin:"auto",width:"500px"}},a.default.createElement("img",{className:"illust-image",src:o.default.image.model.interaction}))),a.default.createElement("div",{className:"part"},"We genetically modify the E.coli, giving them the ability to detect Vibrio concentration, and secrete some compound to supress Vibrio and itself. "),a.default.createElement("div",{className:"part"},"If the AHL concentration exceed a specific threshold, engineered E.coli will secrete antibacterial peptide Colisin M to kill Vibrio. Otherwise, the Vibrio concentration below the E.coli's detection threshold, engineered Ecoli will secrete tsRNA to commit suicide.")),f=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},a.default.createElement("div",null,"We revise this model from Lotka-Volterra model with logistic population groth, and the decay rate of compound."),a.default.createElement("div",{style:{width:"fit-content",margin:"auto"}},a.default.createElement(s.default.Provider,null,a.default.createElement("div",{style:{marginTop:"20px"}},a.default.createElement(s.default.Node,{inline:!0,formula:"{{dV} \\over{dt}} = r_v V - {{r_v} \\over {V_{max}}} V^2 -a VN"})),a.default.createElement("div",{style:{marginTop:"20px"}},a.default.createElement(s.default.Node,{inline:!0,formula:"{{dA} \\over {dt}} = k_AV - \\lambda_A A"})),a.default.createElement("div",{style:{marginTop:"20px"}},a.default.createElement(s.default.Node,{inline:!0,formula:"{{dE} \\over {dt}} = bEA - cES - \\lambda_E E"})),a.default.createElement("div",{style:{marginTop:"20px"}},a.default.createElement(s.default.Node,{inline:!0,formula:"{{dC} \\over {dt}} = k_CE - \\lambda_C C "})),a.default.createElement("div",{style:{marginTop:"20px"}},a.default.createElement(s.default.Node,{inline:!0,formula:"{{dS} \\over {dt}} = k_SE - \\lambda_S S"}))))),a.default.createElement("div",{className:"part"},"With the contraint of AHL threshold",a.default.createElement("div",{style:{width:"fit-content",margin:"auto"}},a.default.createElement(s.default.Provider,null,a.default.createElement("div",{style:{marginTop:"20px"}},a.default.createElement(s.default.Node,{inline:!0,formula:"A \\lt A_t, \\ k_C=0"})),a.default.createElement("div",{style:{marginTop:"20px"}},a.default.createElement(s.default.Node,{inline:!0,formula:"A \\geq A_t, \\ k_S=0"})))))),d=a.default.createElement("div",null,"With changing specific parameter, such as binding strength of antibacterial Colisin with Vibrio, binding strength of Ahl with Ecoli, and the detection threshold of AHL for engineered E.coli. We are able to simulate the population of two bacteria in oscillating, and a parameter sensitive constant mode.",a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.model.result}),a.default.createElement("div",{className:"undertext"},"Two figure are just a slight change of the Binding strength of Ahl with Ecoli"))),p=a.default.createElement("div",null,"Code detail On ",a.default.createElement("a",{href:"https://github.com/igem-tw-nthu/model-2018"},"Github")),h={viewbar:{Title:"Model",Subtitle:"Build a simple bio-oscillator",image:o.default.image.topfull.model},content:{Introduction:"This year 2018 iGEM NTHU_Taiwan aims to solve the Vibrio infection in aquaculture with bio-competiotion. We are inspired by the peroidic pattern of Lotka-Volterra model, and try to control the Vibrio population with engineered E.coli.",parts:[{Title:"Interaction",Description:c},{Title:"Equations",Description:f},{Title:"Discussion",Description:d},{Title:"Soucre Code",Description:p}]}},m=function(e){function t(){return function(e,t){if(!(e instanceof t))throw new TypeError("Cannot call a class as a function")}(this,t),function(e,t){if(!e)throw new ReferenceError("this hasn't been initialised - super() hasn't been called");return!t||"object"!=typeof t&&"function"!=typeof t?e:t}(this,(t.__proto__||Object.getPrototypeOf(t)).apply(this,arguments))}return function(e,t){if("function"!=typeof t&&null!==t)throw new TypeError("Super expression must either be null or a function, not "+typeof t);e.prototype=Object.create(t&&t.prototype,{constructor:{value:e,enumerable:!1,writable:!0,configurable:!0}}),t&&(Object.setPrototypeOf?Object.setPrototypeOf(e,t):e.__proto__=t)}(t,i.Component),r(t,[{key:"render",value:function(){return a.default.createElement(l.default,h)}}]),t}();t.default=m},function(e,t,n){"use strict";Object.defineProperty(t,"__esModule",{value:!0}),t.default=void 0;var r=function(){function e(e,t){for(var n=0;n<t.length;n++){var r=t[n];r.enumerable=r.enumerable||!1,r.configurable=!0,"value"in r&&(r.writable=!0),Object.defineProperty(e,r.key,r)}}return function(t,n,r){return n&&e(t.prototype,n),r&&e(t,r),t}}(),i=n(0),a=s(i),o=s(n(4)),l=s(n(8));function s(e){return e&&e.__esModule?e:{default:e}}var u=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Transformation"),a.default.createElement("div",null,"During this summer, we had done lots of digestion-ligation and transformation experiments! We enjoyed and had fun exploring the mystery of synthetic biology!"),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.experiment.transform}),a.default.createElement("div",{className:"undertext"},"Fig. 1: Successful transformation of part BBa_K2774008 (Left) and BBa_K2774005 (Right)")),a.default.createElement("div",null,"Utilizing iGEM distribution kit and parts from the registry, we assembled various bio-bricks to construct our genes. We constructed our part, BBa_K2774008, by using pLac with RBS (BBa_J04500), endolysin (BBa_K1378032) with GFP (E0240). The left LB plate was the transformation of BBa_K2774008. The transformation was success and therefore some of the colony emits GFP fluorescence! And the right LB plate was the transformation of part BBa_K2774005, the RFP shows the right colony with successful ligation plasmid. In addition, thanks for the established technology of IDT (Integrated DNA Technologies), providing us the gene fragments. Finally, we successfully constructed tsRNA (BBa_K2774001).")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Electrophoretogram"),a.default.createElement("div",null,"Here are the parts we assemble successfully. We use VF2 and VR primer to check our gene’s length, all of them are on pSB1C3."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.experiment.elec}),a.default.createElement("div",{className:"undertext"},"Fig.2: Electrophoretogram of our parts which we had submitted to registry, all of them show properly length (Using VF2 and VR primer, the length will increase about 300 bp)")),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.experiment.table_1}),a.default.createElement("div",{className:"undertext"},"Table 1: This table’s genes correspond to each lane of Fig. 2 from left to right.")))),c=a.default.createElement("div",null,"We use TCBS (Thiosulfate-citrate-bile salts-sucrose agar, a type of selective agar culture plate that is used in microbiology laboratories to isolate Vibrio) to examine Vibrio’s concentration in fish farm. Our HP member and wet lab member visit the fish farm in Taiwan, Chiayi. We collected the water sample from different pool, and sent it back to the lab to culture the sample on TCBS.",a.default.createElement("div",{className:"illustration"},a.default.createElement("div",{className:"illust-image-wrapper",style:{width:"600px"}},a.default.createElement("img",{className:"illust-image",src:o.default.image.experiment.sample})),a.default.createElement("div",{className:"undertext"},"Fig.3 : Collection sample from traditional fish farm in Taiwan")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Result"),a.default.createElement("div",null,"Sample from fish farm shown positive result, we found there were colony appeared on TCBS plate. Implying that Vibrio indeed exist in fish farm, and may infect shrimp or other marine creatures. In addition, the colony color stands for different types of Vibrio:"),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",null,"Vibrio Cholerae: Large yellow colonies. "),a.default.createElement("div",null,"Vibrio harveyi / Vibrio fischeri: Greyish-green to bluish-green colonies which show luminescence in dark.")),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:o.default.image.experiment.vibrio}),a.default.createElement("div",{className:"undertext"},"Fig.4 : Testing Sample from fish farm on TCBS"),a.default.createElement("div",{className:"undertext"},"Right plate shows Vibrio harveyi and the left pate shows Vibrio Cholerae")))),f=a.default.createElement("div",null,"Our detection system is designed to make E. coli to obtain the ability to sense the concentration of Vibrio in the environment. We found that most of the pathogenic Vibrio use “quorum sensing” to communicate with each individual and to express different genes. Quorum sensing, a system which is able to control the gene expression followed by the population density. The signal transmission of quorum sensing is totally depending on the auto-inducer, acyl homoserine lactones (AHL), a small chemical molecule which is produced by bacterial enzyme (Fig. 5). Moreover, the concentration of AHL is directly proportional to bacteria density. When AHL concentration exceeds the threshold, it will bind with the protein (LuxR) and trigger the promoter (pLUX) to express the downstream genes.",a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:o.default.image.experiment.ahl}),a.default.createElement("div",{className:"undertext"},"Fig. 5: The schematic of our gene circuits")),a.default.createElement("div",{className:"illustration",style:{width:"300px"}},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:o.default.image.experiment.structure}),a.default.createElement("div",{className:"undertext"},"Fig. 6: The structure of AHL")),"Our detection system is using Lux system. When the Vibrio density is high, the AHL concentration in the environment is high. Hence, our engineered E. coli can detect the Vibrio density and can release the anti-Vibrio substances. (Fig. 7)",a.default.createElement("div",{className:"illustration"},a.default.createElement("div",{className:"illust-image-wrapper",style:{width:"500px"}},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:o.default.image.experiment.cycle})),a.default.createElement("div",{className:"undertext"},"Fig. 7: The schematic of our detection system")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Exp. 1: AHL Sensor with Quantitative AHL "),a.default.createElement("div",null,"We constructed our AHL detector (BBa_K2774000), which can produce LuxR consistently by pLac promoter and can express GFP while LuxR combine with AHL and trigger pLUX (R0062). While AHL presents, AHL and LuxR will form a complex and trigger pLUX to express GFP. (Fig. 8)"),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.experiment.sensor}),a.default.createElement("div",{className:"undertext"},"Fig. 8: AHL Bio-Detector")),a.default.createElement("div",null,"In order to test our gene’s function, we added different concentration of AHL (3-oxo-C6-HSL, synthesized by Sigma) to DH5α cells transformed with BBa_K2774000, and measured GFP expression hourly."),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Procedure:"),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",{className:"innerpart"},"1. Prepare DH5α cells which transformed with BBa_K2774000 and liquid culture in LB overnight."),a.default.createElement("div",{className:"innerpart"},"2. Diluted eight bacterial culture samples in fresh LB (50 μL O/N culture with 4950 μL fresh LB)"),a.default.createElement("div",{className:"innerpart"},"3. Pre-culture until O.D.600 reached 0.4, making sure bacteria is growing in log phase (better expression capability) and add IPTG (1mM) to induce the expression of LuxR."),a.default.createElement("div",{className:"innerpart"},"4. Add different amount of AHL into different samples (0.01, 0.1, 0.5, 1, 5, 10, 100 nM) and 0 M AHL for positive control."),a.default.createElement("div",{className:"innerpart"},"5. Measure the O.D.600 value and expression of GFP (λexcitation = 490 nm; λemission = 530 nm) hourly.")),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.experiment.exp_1}),a.default.createElement("div",{className:"undertext"},"Fig. 9: GFP expression from different conc. of AHL"))),a.default.createElement("div",{className:"part"}),a.default.createElement("div",{className:"part-title"},"Result:"),a.default.createElement("div",null,"As the chart shows, we can find that AHL induced GFP expression as we expected. As the AHL concentration increased, the GFP expression increased until saturation. Besides, the optimized AHL concentration of inducing BBa_K2774000 is about 0.5-10 nM. Moreover, we found that if the concentration of AHL exceeds 1000 nM, it will inhibit BBa_K2774000 GFP expression."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:o.default.image.experiment.data_1}),a.default.createElement("div",{className:"undertext"},"Fig. 10: GFP expression from different conc. of AHL"))),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Exp. 2 : AHL sensor co-culture with fake Vibrio test"),"Since we cannot conduct the experiment related to Vibrio in our lab, we have to construct a fake Vibrio (by our engineered E. coli), which can also secrete AHL as Vibrio does. Therefore, we created our “pLac AHL generator” (BBa_2774004) by assembling “pLac + B0034” (BBa_J04500) and “LuxI” (BBa_C0161) together. Moreover, we added RFP to detect the population of fake vibrio (BBa_2774005)",a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Procedure:"),a.default.createElement("div",{className:"innerpart"}," 1. Prepare DH5α cells which transformed with BBa_2774000 (AHL sensor), BBa_2774004 (pLac AHL generator), and BBa_2774005 (pLac AHL generator + RFP) and liquid culture in LB overnight."),a.default.createElement("div",{className:"innerpart"}," 2. Diluted eight overnight bacterial culture in fresh LB with 1mM IPTG (50μL O/N culture with 4950 μL fresh LB)."),a.default.createElement("div",{className:"innerpart"}," 3. Pre-culture until O.D.600 reaches 0.4, making sure bacteria is growing in log phase (better expression capability)."),a.default.createElement("div",{className:"innerpart"}," 4. Mix different volume ratio of:"),a.default.createElement("div",null," BBa_2774004 (pLac AHL generator) with BBa_2774000 (AHL sensor)    "),a.default.createElement("div",null," BBa_2774005 (pLac AHL generator + RFP) with BBa_2774000 (AHL sensor)"),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:o.default.image.experiment.table_2})),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:o.default.image.experiment.table_3})),a.default.createElement("div",null,"5. Measure O.D.600, GFP and RFP expression after overnight culture.")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Result:"),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"a. BBa_2774005 (pLac AHL generator + RFP) co-culturing with BBa_2774000 (AHL sensor)"),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:o.default.image.experiment.exp_2}),a.default.createElement("div",{className:"undertext"},"Fig 11. AHL sensor with increasing concentration of AHL generator + RFP (from the left tube to the right tube). We centrifuged culture tubes in the left picture to get the ones in the right picture.")),a.default.createElement("div",{className:"innerpart"},"We co-cultured AHL sensor with increasing concentration of AHL generator + RFP. The picture indicates that AHL was indeed generated. When the two bacteria were mixed together, red and green fluorescence were both emitted and producing different colors.")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"b. BBa_2774004 (pLac AHL generator) co-culturing with BBa_2774000 (AHL sensor) "),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:o.default.image.experiment.table_4})),a.default.createElement("div",{className:"innerpart"},"In this experiment, we co-cultured AHL sensor with increasing concentration of AHL generator. We discovered that all liquid culture emitted green fluorescence except the one without adding AHL generator. We successfully proved that BBa_2774004 can express LuxI (AHL synthase) and generate AHL. We proposed that AHL concentration is linearly proportional to fake Vibrio concentration. However, the fluorescence intensities were almost the same among samples with increasing concentrations of AHL. We failed to construct a rational relation between fake Vibrio density and AHL concentration. We inferred the reason for this observation is that the rate of AHL synthesis is much higher than the rate of AHL degradation. The LuxI we used was the one without LVA-tag, so LuxI didn’t degrade instantly. In addition, we left the liquid culture overnight before measuring its fluorescence. Consequently, AHL was over-generated and the concentration of AHL was beyond the detection limit of AHL sensor.")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Exp. 3: Culture AHL sensor with Fake Vibrio’s supernatant")),a.default.createElement("div",null,"During Exp. 2 we find that overnight culturing and strong promoter cause excess AHL, which will not show the evidence of AHL concentration proportion to fake Vibrio concentration. Therefore we try a different way to experiment, we use weaker promoter and measure AHL concentration by AHL bio-sensor every hourly. Using interlab data we can get bacteria concentration by measuring OD600, and meanwhile we can centrifuge the tube to get supernatant which contain AHL secrete by the AHL generator that time."),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Procedure:"),a.default.createElement("div",{className:"innerpart"},"1. Prepare DH5α cells which transformed with BBa_2774000 (AHL sensor), BBa_2774002 (J23119+LxuI), and liquid culture in LB overnight."),a.default.createElement("div",{className:"innerpart"},"2.  Culture BBa_2774002 at microcentrifuge tube with (50μL O/N culture with 4950 μL fresh LB)."),a.default.createElement("div",{className:"innerpart"},"3. Measure microcentrifuge’s OD600 two hour and meanwhile centrifuge the tube to get supernatant contain AHL. Transfer the OD600 data to bacteria density by using inter lab data."),a.default.createElement("div",{className:"innerpart"},"4. Move 2.5 ml of supernatant contain AHL and 2.5ml fresh LB in new tube, and add AHL sensor simultaneously."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:o.default.image.experiment.exp_3}),a.default.createElement("div",{className:"undertext"},"Fig 12. The procedure of experiment 3."))),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Result:"),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:o.default.image.experiment.result_4}),a.default.createElement("div",{className:"undertext"},"Fig 13. AHL sensor with increasing concentration of AHL generator supernant")),a.default.createElement("div",{className:"innerpart"},"We transferred the supernatant from generator liquid culture which contained AHL into AHL sensor. The result showed that supernatant from a higher concentration of generator inducing AHL sensor to express higher amount of GFP. Moreover, after we transform the data and take the logarithm of generator density, we find a similar result which we had done in experiment 1. (Fig10)"),a.default.createElement("div",{className:"innerpart"},"Comparing exp1 and exp3 result, confirming that Vibrio density is proportional to the AHL concentration, at the condition of the real-time situation to avoid AHL accumulation. ")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Conclusion:"),a.default.createElement("div",null,"We have successfully created a bio-sensor which can detect and react to different concentration of AHL. Moreover, our fake Vibrio (BBa_2774004) can also secrete AHL. We also confirmed that the Vibrio density is proportional to the AHL concentration. Our future work is to exchange gene which is under the control of pLux from GFP to anti-Vibrio substances."))))),d=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"innerpart"},"Based on our gene circuit, we need two kinds of substances to fulfill our goal. The first one is the suicide gene. When Vibrio concentration is low, our E. coli need to suicide themselves to maintain the equilibrium. Therefore, we introduced “tsRNA” (toxic small RNA), which was studied and experimented by Dr. Rotem . “tsRNA” was first found in Burkholderia cenocepacia and showed its toxic to E. coli. to inhibit E. coli growth. Moreover, references show that “tsRNA” couldn’t penetrate the bacterial cell wall, either from outside or inside. Therefore, it becomes the best candidate for our suicide gene. Although the mechanism of “tsRNA” is not clearly known, Dr. Rotem had tried complement sequences of “tsRNA” shown negative results. They hypothesized that “tsRNA” might target and mis-regulate essential genes within the E. coli cell. "),a.default.createElement("div",{className:"innerpart"},"To test the function of tsRNA for our suicide gene, we put tsRNA sequence under pLac promoter. Using IDT synthesized technology, we got the gene fragment (BBa_2774001)."),a.default.createElement("div",{style:{width:"300px"},className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:o.default.image.experiment.tsrna}),a.default.createElement("div",{className:"undertext"},"Fig 14. The secondary structure of tsRNA"))),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Procedure"),a.default.createElement("div",{className:"innerpart"},"1. Prepare DH5α which transformed with BBa_K2774001 and liquid culture in LB overnight."),a.default.createElement("div",{className:"innerpart"},"2. Diluted the overnight bacterial culture in fresh LB (50 μL O/N culture with 4950 μL fresh LB) "),a.default.createElement("div",{className:"innerpart"},"3. Pre-culture until O.D.600 reaches 0.2, making sure bacteria is growing in log phase (better expression capability) "),a.default.createElement("div",{className:"innerpart"},"4. Add IPTG (1mM) to induce tsRNA expression. (Adding ampicillin for positive control)"),a.default.createElement("div",{className:"innerpart"},"5. Measure O.D.600 hourly."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:o.default.image.experiment.tsrna_test}))),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Result"),a.default.createElement("div",{className:"innerpart"},"We induced tsRNA expression with IPTG at O.D.600 = 0.2. The result shows that tsRNA slightly inhibits E. coli growth, approximately 12%. We tested our BBa_K2774001 lots of times, but it still goes to the same result. Feeling confused, so we consulted Dr. Rotem for our situation, and got a generous reply:"),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:o.default.image.experiment.email})),a.default.createElement("div",{className:"innerpart"},"Dr. Rotem suggested us to check if our tsRNA mutated during transforming because of the inferiority of growth rate. Or to check tsRNA expression by using Northern blotting to check the RNA expression. Therefore, we sequenced our plasmid (tsRNA) and BLAST the result with our designed sequence: showing no mutation on tsRNA section."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:o.default.image.experiment.strength}),a.default.createElement("div",{className:"undertext"},"Fig 15. Sequencing result of tsRNA")),a.default.createElement("div",{className:"part-title"},"Conclusion"),a.default.createElement("div",{className:"innerpart"},"Our experiment result shows that the inhibition efficiency of tsRNA is 12%, which is significantly different from the one reported by Dr. Rotem. We sequenced our plasmid and no mutation was found. We inferred that the low inhibition efficiency is due to few expression of tsRNA. We believe this can be improved by using a stronger promoter to get a better result."))),p={viewbar:{Title:"Experiment",Subtitle:"",image:o.default.image.topfull.experiment},content:{Introduction:"In order to achieve our project’s goal, we constructed a few functional genes and tested their features. To sum up, our project can be divided into three parts. First of all is the “detection” part, we made our E. coli to detect the density of Vibrio and to respond. Then, it’s the part of “kill Switch”, we made our E. coli to switch from secreting anti-Vibrio substances to suicide to fulfilled bio-safety issues. Finally, it’s  “substances test” for anti-Vibrio and suicide substances which will be used in our project.",parts:[{Title:"Cloning",Description:u},{Title:"Testing Sample",Description:c},{Title:"Detection",Description:f},{Title:"Substances test",Description:d}]}},h=function(e){function t(e){return function(e,t){if(!(e instanceof t))throw new TypeError("Cannot call a class as a function")}(this,t),function(e,t){if(!e)throw new ReferenceError("this hasn't been initialised - super() hasn't been called");return!t||"object"!=typeof t&&"function"!=typeof t?e:t}(this,(t.__proto__||Object.getPrototypeOf(t)).call(this,e))}return function(e,t){if("function"!=typeof t&&null!==t)throw new TypeError("Super expression must either be null or a function, not "+typeof t);e.prototype=Object.create(t&&t.prototype,{constructor:{value:e,enumerable:!1,writable:!0,configurable:!0}}),t&&(Object.setPrototypeOf?Object.setPrototypeOf(e,t):e.__proto__=t)}(t,i.Component),r(t,[{key:"render",value:function(){return 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We both aimed to let students know more about iGEM and synthetic biology. During the camp, participants learned how to carry out simple experiments, e.g. plasmid extraction and transformation. We also shared with the students about our iGEM project and briefly introduced synthetic biology. The following are the schedule and all the material we used in the camp. About our files used in this camp are all available to",a.default.createElement("a",{href:"https://drive.google.com/drive/folders/19QhZWbbpJTddxGISAeXFJBAxcs_MuS_m?usp=sharing"}," download"),". The only thing you have to notice is the language, most are in Chinese."),f=a.default.createElement("div",null,a.default.createElement("div",{className:"illustration"},a.default.createElement("table",{className:"table"},a.default.createElement("tbody",null,a.default.createElement("tr",{className:"tablehead"},a.default.createElement("th",{className:"tablehead"}),a.default.createElement("th",{className:"tablehead"},"2018/08/10"),a.default.createElement("th",{className:"tablehead"},"2018/08/11")),a.default.createElement("tr",{className:"tablehead"},a.default.createElement("th",{className:"tablehead"},"9:00~12:00"),a.default.createElement("td",{className:"tabledata"},a.default.createElement("div",null,"iGEM Introduction & Team Project"),a.default.createElement("div",null,"Experiment Introduction "),a.default.createElement("div",null,"Pipetman Teaching ")),a.default.createElement("td",{className:"tabledata"},a.default.createElement("div",null,"Synthetic Biology Introduction"),a.default.createElement("div",null,"Experiment Introduction"),a.default.createElement("div",null,"Colony PCR"))),a.default.createElement("tr",{className:"tablehead"},a.default.createElement("th",{className:"tablehead"},"12:00~13:00"),a.default.createElement("td",{className:"tabledata"},"Lunch Time"),a.default.createElement("td",{className:"tabledata"},"Lunch Time")),a.default.createElement("tr",{className:"tablehead"},a.default.createElement("th",{className:"tablehead"},"13:00~16:00"),a.default.createElement("td",{className:"tabledata"},a.default.createElement("div",null,"Plasmid Extraction"),a.default.createElement("div",null,"Transformation"),a.default.createElement("div",null,"Bacteria Cultivation")),a.default.createElement("td",{className:"tabledata"},a.default.createElement("div",null,"Colony PCR"),a.default.createElement("div",null,"DNA Electrophoresis"),a.default.createElement("div",null,"Questionnaire Survey"))))))),d=a.default.createElement("div",null,a.default.createElement("div",null,"This year, 16 students joined our synthetic biology camp. The following are some feedback given by the students in the questionnaire survey. 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Or the students would be boring."),a.default.createElement("tr",{className:"tablehead"},a.default.createElement("td",{className:"tabledata"},"The contents of the experiments were quite funny, but the basic theories were difficult for high school students.")),a.default.createElement("tr",{className:"tablehead"},a.default.createElement("td",{className:"tabledata"},"Though I have not learned most of the terms about molecular biology experiments, the contents of experiments were interesting and inspiring which makes me acquire knowledge about molecular biology.")),a.default.createElement("tr",{className:"tablehead"},a.default.createElement("td",{className:"tabledata"},"If lecturers summarized all of the experiments to a table, it will be better!")),a.default.createElement("tr",{className:"tablehead"},a.default.createElement("td",{className:"tabledata"},"It’s my first time to do the molecular biology experiments and hopefully, you guys can achieve your goals!"))))))),p=a.default.createElement("div",null,a.default.createElement("div",null,"According to their feedback, we have achieved our goal: First, to introduce iGEM and synthetic biology. Second, teach high school students how to carry out synthetic biology experiment. Third, enhance their interest in genetic engineering. The synthetic biology camp this year is successful, and National Feng-Shan Senior High School are willing to cooperate with us next year! We hope our effort can encourage more students to do researches about genetic engineering, and maybe they will join iGEM soon!"),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:l.default.image.education.group}))),h={viewbar:{Title:"Education",Subtitle:u,image:l.default.image.topfull.education},content:{Introduction:"Joining iGEM allows us to carry out some interesting and creative experiments related to the latest researches. However, for ordinary senior high school students in Taiwan, they do not have the opportunity to engage in iGEM or the state-of-the-art knowledge of synthetic biology. Therefore, we decided to bring synthetic biology to high school students. We can not only tell them what synthetic biology is but also share our experience of joining iGEM as well as encourage them to participate next year!",parts:[{Title:"Summer Synthetic Biology Camp",Description:c},{Title:"Schedule",Description:f},{Title:"Feedback",Description:d},{Title:"Summary",Description:p}]}},m=function(e){function t(e){return function(e,t){if(!(e instanceof t))throw new TypeError("Cannot call a class as a function")}(this,t),function(e,t){if(!e)throw new ReferenceError("this hasn't been initialised - super() hasn't been called");return!t||"object"!=typeof t&&"function"!=typeof t?e:t}(this,(t.__proto__||Object.getPrototypeOf(t)).call(this,e))}return function(e,t){if("function"!=typeof t&&null!==t)throw new TypeError("Super expression must either be null or a function, not "+typeof t);e.prototype=Object.create(t&&t.prototype,{constructor:{value:e,enumerable:!1,writable:!0,configurable:!0}}),t&&(Object.setPrototypeOf?Object.setPrototypeOf(e,t):e.__proto__=t)}(t,i.Component),r(t,[{key:"render",value:function(){return a.default.createElement(o.default,h)}}]),t}();t.default=m},function(e,t,n){"use strict";Object.defineProperty(t,"__esModule",{value:!0});var r=function(){function e(e,t){for(var n=0;n<t.length;n++){var r=t[n];r.enumerable=r.enumerable||!1,r.configurable=!0,"value"in r&&(r.writable=!0),Object.defineProperty(e,r.key,r)}}return function(t,n,r){return n&&e(t.prototype,n),r&&e(t,r),t}}(),i=n(0),a=s(i),o=(s(n(10)),s(n(4))),l=s(n(8));function s(e){return e&&e.__esModule?e:{default:e}}var u=a.default.createElement("div",null,a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.interview.research})),a.default.createElement("div",null,"Fisheries Research Institute, COA, which is the government agency offers fish farmers to test the pathogen of fish and also, doing some researches on aquatic organisms as well. Deputy director of COA, C.I. Chang, who is a master in marine biology, recently did lots of researches on aquatic animal and pathogen, creating the MV-kit as well. The kit which function is to test the number of Vibrio in the water. And of course, FRI is the closest department between the local fish farmers and the public. As a result, we decided to visit him to get more helpful information and suggestions for our project. During our in-depth talk, we knew more about the Vibrio and, our core of the automated device,",a.default.createElement("a",{href:"https://2018.igem.org/Team:NTHU_Taiwan/Drylab-Overview"}," ACE"),". For example, after knowing the detailed information about the basic characteristics of Vibrio and the mechanism of MV-kit, making us want to combine the kit with our device and if it comes true, the device will be powerful. In the end, deputy director also gave us a few useful, potential advice and questions between our device and the kit. These feedbacks inspired us pretty much, such as whether our device would be influenced by the precipitation or not, or do we need some kinds of membrane to filter the aquatic sample or not, and in order to maintain the integrity and accuracy of our device, we have to test the efficiency of MV-kit in different environments and to find the simplest way that fish farmers can easily use. Nicely, the deputy director offering the plate of Vibrio harveyi and MV-kit for free in order to encourage us to make our experiment more complete.")),c=a.default.createElement("div",null,"In order to improve our project, we need to gain as much feedback as possible from the potential users; therefore, we’ve visited local fisherman in Taiwan, of which we can categorize into ecological and traditional breeding fish farmers."),f=a.default.createElement("div",null,a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.interview.tradition_1})),a.default.createElement("div",null,a.default.createElement("div",null,"In addition to visiting the ecological breeding farmers, we visited traditional fishermen in Kaohsiung and Chiayi, Mr. Chiu and Mr. Jian. They gave us some useful information, so we can improve our project more further."),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Mr. Chiu and Mr. Jian"),a.default.createElement("div",{className:"innerpart"},"Overall, they usually wait until the fish get sick and take them to the inspection center. If we can let them know about the condition of bacteria beforehand, they might take precautions, such as cleaning the water or adding the active powder to improve water quality and minimize the damage."),a.default.createElement("div",{className:"innerpart"},"When asking about what problem is the most crucial one they are facing, they’ve mentioned the uncertainty of electricity.  If the machine shut down or in a dangerous condition, sending a notification to farmers is essential. They also mentioned that adding the function of notifying farmer by message might be more practical than the smartphone application or the notification bar since farmer might be the elderly who do not use a smartphone. In light of their suggestions, we found the Chunghwa Telecom to seek for future collaboration. They told us that as long as we apply for a SIM card and a phone number, this kind of function can easily be done."),a.default.createElement("div",{className:"innerpart"},"To sum up, feedbacks and information are listed below. Plenty of them has greatly improved our projects:"),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",null,"1. Their working time is usually between 2 a.m. to 11 a.m., so they suggest us to test the sample during this time (since MV-kit needs 12~24 hr. to react completely)."),a.default.createElement("div",null,"2. They introduce the “TCBS” method: a common way to detect Vibrio sp. in traditional fish farm. This method need 1~2 day to complete."),a.default.createElement("div",null,"3. Add more features to ACE machine, e.g. water quality test (pH, NO3-, NO2-), other kit test (virus, fish parasite), and black out reminder."),a.default.createElement("div",null,"4. They recommend us to use text messages to be the notification of ACE machine. Because some of the traditional fishermen do not use smartphones."),a.default.createElement("div",null,"5. They recommend us to cooperate with fishermen who feed grouper or eel."),a.default.createElement("div",null,"6. As a traditional fisherman, they think our device has great potential."))))),d=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Mr. Tsai Hung-Jing’s Fish Farm"),a.default.createElement("div",null,a.default.createElement("div",{className:"innerpart"},"Before deciding which fish farm to visit, we looked for information on the Internet and finally found an interview published by CommonWealth Magazine, which is a famous magazine in Taiwan. The article showed that how successfully Mr. Tsai uses biological feed - water flea to take care of his shrimps and to make some profit. This idea of substituting chemical feed into water flea, an eco-friendlier one, matched the concept of iGEM. So, we called him and arranged the visit time without hesitation."),a.default.createElement("div",{className:"innerpart"},"Mr. Tsai was nice and friendly, he not only introduced his fish farm in such a detailed way but also answered all of our questions. He mentioned that the way he used to prevent Vibrio is to make the environment beneficial for good germs, which occupies the environment of the Vibrio, leaving no spaces for them to grow. He pointed out that people often uses ClO2 to kill Vibrio is unable to eradicate the problem completely. Once the pollutants are in, Vibrio will grow again. Consequently, we introduced our method to him and he sounded satisfied and looked forward to our final result. He also gave us some positive feedback toward our ACE. He said that if our service cost can be as low as 8,000 NT dollars, it will have a strong advantage and the market acceptance will be high since the price of the general detection devices need to be up to ten thousand NT dollars."),a.default.createElement("div",{className:"innerpart"},"The interview with Mr. Tsai really cheered us up, he let us understand the thing we do might help them in a practical way. Even though we went back to school, he still sent messages to encourage us.")),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.interview.eco_1}))),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Qiujia Aquaculture in Chiayi County"),a.default.createElement("div",null,a.default.createElement("div",{className:"innerpart"},"Qiujia Aquaculture has been established for 50 years, the main breed they sell are milkfish and shrimps. The reason why we wanted to visit them was that they refuse to use any of the chemical agents or drug and it triggered our curiosity, how do they make it?"),a.default.createElement("div",{className:"innerpart"},"By actually visiting them on the ground, we discovered that they do not use any chemical medicine or drug to kill bacteria, instead, they use Bacillus subtilis to compete with Vibrio. We surprisingly found out this kind of ecological competence is similar to what our wet lab is trying to mimic. Though adding good germs can’t guarantee no fish will be infected, they believed doing so was far better than adding artificial chemicals which have huge uncertainty to humankind. When asking about whether they would detect the concentration of Vibrio, they told us the current method they use to detect Vibrio is coating the plate, which is not efficient and not so accurate."),a.default.createElement("div",{className:"innerpart"},"Furthermore, they provided us with pieces of advice and feedback regarding our Automatic Color-Detecting Examiner (ACE):"),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",null,"1.ACE needs to be equipped with high accuracy, shutdown notification, and self-calibration."),a.default.createElement("div",null,"2. If they knew the concentration of Vibrio earlier, they can take actions to prevent the Vibrio from growing too much beforehand, such as adding Bacillus subtilis."),a.default.createElement("div",null,"3. Since the growing time of Vibrio to be pathogenic is way longer than one day, a period of sampling doesn’t need to be as frequent as 1 hour, but one day. "),a.default.createElement("div",null,"4. They suggested us to target indoor breeding customers, since that if the device is set outdoors, it may be easily broken down. Moreover, fish with high economic value bred indoor might be more fragile than the outdoors.")),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.interview.eco_2})),a.default.createElement("div",null,"After considering their suggestions, we decided to extend the sampling period to 12 hours, adding the fool-proof design, such as that if the camera shut down or cannot be connected, it will return a “no camera” sign to our application.")))),p=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},a.default.createElement("div",null,a.default.createElement("div",{className:"part-title"},a.default.createElement("a",{href:"http://dataweb.nmmba.gov.tw/ResearchMemberDetail.aspx?CondID=bfcdc17d-da5f-48f8-9af8-819a3376101a"},"Dr. Jimmy Kuo")),"Dr. Jimmy Guo, a vice researcher of the national museum of marine biology and aquarium, have done many types of research of Vibrio in the aquarium. He first shared with us how he made the research on Vibrio and explained why the growth curve of Vibrio was so strange. The reason is that Vibrio is a special species which is different from other normal bacteria. Vibrio will be dormant and make its metabolism disordered when stored at low temperature. Therefore, if we want to get the expectative results, we should culture fresh Vibrio rather than take one from the refrigerator."),a.default.createElement("div",null,"Besides the advice for the wet lab, Dr. Guo also gave us some suggestions to our hardware. He recommended that we could use the battery as our electric power source and thus, avoiding the loss of results from the blackout. Additionally, our device should calibrate regularly, or the results would be deviant. Finally, since our device could detect frequently, we could target on the customers cultivating remunerative aquatic agriculture."),a.default.createElement("div",null,a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.interview.aqua_1})))),a.default.createElement("div",{className:"part"},a.default.createElement("div",null,a.default.createElement("div",{className:"part-title"},a.default.createElement("a",{href:"https://www.nmmba.gov.tw/research/RMenber_Content.aspx?n=291A05EEDDA1D4C4&sms=CF7FFA69EB91FF61&s=6A6B57F5FE966020"},"Dr. Ming-An Tsai ")),"Dr. Ming-An Tsai, an assistant researcher in the national museum of marine biology and aquarium, is interested in doing research on diseases of aquatic animals. Fortunately, Dr. Tsai’s lab is also doing some Vibrio-detecting experiment, so we first exchanged some ideas about Vibrio experiments. After introducing our project, Dr. Tsai gave us some important advice. First of all, he suggested us that we should ensure which way we would use to avoid bacterial diseases, killing Vibrio or inhibiting its growth. Additionally, he reminded us that we should not kill all the Vibrio in the water, or we might destroy the ecological balance. Finally, he gave us a crucial reference that if the concentration of Vibrio in the water goes beyond 104 CFU/mL, it would be more infectious. This data is a vital reference not only in our wet lab but also in our dry lab."),a.default.createElement("div",null,a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.interview.aqua_2}))))),h={viewbar:{Title:"Interview",Subtitle:"Visited the research institute, fishing farm, and aquarium",image:o.default.image.topfull.interview},content:{Introduction:"Our project is an interdisciplinary work which is so comprehensive and complete to makes it possesses a potent potential to practice in the real world. Based on this advantage, to navigate our project toward a suitable direction to either meet the true demand and solve real problem is quite essential. At the beginning, we visited Deputy director of Fisheries Research Institute. He gave us some advice on how to start our project. Then, we had conducted interviews with fishermen in Taiwan, visited the fishing farm with them and received countless useful information about how to alter our design to fit their demand and improve the latest solution they adopt to combat bacterial infection. Moreover, with the advice given by researchers from national museum of marine biology and aquarium, we gained much more practical perspective on what’s the pros and cons of our solution and how we can lower the risk that will harm the environment. With their help, we have achieved the core value of human practice: know who need, find what they need and meet their need.        ",parts:[{Title:"Fisheries Research Institute, COA ",Description:u},{Title:"Interview with Fish Farmer",Description:c},{Title:"Ecological Breeding",Description:d},{Title:"Traditional Fishermen",Description:f},{Title:"National Museum of Marine Biology and Aquarium",Description:p}]}},m=function(e){function t(){return function(e,t){if(!(e instanceof t))throw new TypeError("Cannot call a class as a function")}(this,t),function(e,t){if(!e)throw new ReferenceError("this hasn't been initialised - super() hasn't been called");return!t||"object"!=typeof t&&"function"!=typeof t?e:t}(this,(t.__proto__||Object.getPrototypeOf(t)).apply(this,arguments))}return function(e,t){if("function"!=typeof t&&null!==t)throw new TypeError("Super expression must either be null or a function, not "+typeof t);e.prototype=Object.create(t&&t.prototype,{constructor:{value:e,enumerable:!1,writable:!0,configurable:!0}}),t&&(Object.setPrototypeOf?Object.setPrototypeOf(e,t):e.__proto__=t)}(t,i.Component),r(t,[{key:"render",value:function(){return a.default.createElement(l.default,h)}}]),t}();t.default=m},function(e,t,n){"use strict";Object.defineProperty(t,"__esModule",{value:!0}),t.default=void 0;var r=function(){function e(e,t){for(var n=0;n<t.length;n++){var r=t[n];r.enumerable=r.enumerable||!1,r.configurable=!0,"value"in r&&(r.writable=!0),Object.defineProperty(e,r.key,r)}}return function(t,n,r){return n&&e(t.prototype,n),r&&e(t,r),t}}(),i=n(0),a=s(i),o=(s(n(10)),s(n(4))),l=s(n(8));function s(e){return e&&e.__esModule?e:{default:e}}var u=a.default.createElement("div",null,a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.public.nctu})),a.default.createElement("div",null,"Engaging with other iGEM teams to exchange ideas and to brainstorm together is a crucial part of iGEM. Therefore, we joined a half-day meet-up with three iGEM teams in Taiwan, NTHU_Formosa, NCTU_Formosa, and Mingdao. During the meet-up, we presented our projects, had some discussions with other teams, and received some problems and feedbacks on our projects. After brainstorming, we knew what the problem was and what we could do to improve the project. This meet-up truly broadened our horizons and we got loads out of it. Following are some feedbacks we received from the meet-up:"),a.default.createElement("div",null,a.default.createElement("div",{className:"part"}," 1. There were some drawbacks of our project of biosafety if we put our engineered E. coli into the open water area. After discussion, we finally decided to replace this way by putting it into a close water and to use it with a filter. By doing so, we could effectively control our engineered bacteria and thus, reducing the problem about biosafety. "),a.default.createElement("div",{className:"part"}," 2. We should ensure our device can really benefit the users. Therefore, we visited some local fishermen in person and sought for their suggestions. To find out more, check the ",a.default.createElement("a",{href:"https://2018.igem.org/Team:NTHU_Taiwan/Interview"},"interview")," page"))),c=a.default.createElement("div",null,a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.public.meetup_group})),a.default.createElement("div",null,"In late July, we participated in the first Eurasian meet-up held by BGI college and TU Delft in Shenzhen, China. There are 20 stunning teams from 5 different countries visited Shenzhen, discussing and sharing all kinds of iGEM issues and projects during the conference. Also, we presented and conveyed our idea to other teams, which lets us receive several feedbacks and they were quite useful for our project. At this point, we did find some inspiring ideas which sharpen our current work. Here we conclude something that influenced our work:"),a.default.createElement("div",null,a.default.createElement("div",{className:"part"},"1. The other iGEMers provided us with some useful feedbacks, for instance, they suggested us to integrate and double-check the consistency of data from our bio-reactor and Vibrio-detecting system. Also, team members from CUHK gave us some suggestions about our image detection system. Moreover, some people suggested us other switch systems for our experimental design like toehold switch. After the conference, we searched for several switch systems but we still stick to our original idea because we thought that the disadvantage for our STAR system is the least."),a.default.createElement("div",{className:"part"},"2. A more concise presentation should be considered. This is the first presentation after our frame was formed out, but we spent much time explaining the detailed information about our project. Furthermore, we didn’t integrate our wet lab works and dry lab works in our presentation which was not convincing to the audience. After the presentation, we then modified the whole slides into a more concise and more comprehensible version which do help more audiences to get into our story."),a.default.createElement("div",{className:"part"},"3. The meet-up official set up several workshops and lectures during the conference. For instance, they invited former iGEM judge and iGEM ambassador to share their experiences like “how to be an iGEMer?” from many different perspectives. It seems to us that we learned more details and tips about how to handle the iGEM project and figured out the overview of this competition.")),a.default.createElement("div",null,"Last but not least, except for the feedback we received, we also build a strong friendship connection there. The meet-up official assigns a buddy team for each team and our buddy is Marburg university from Germany. We really enjoyed the moment with them. "),a.default.createElement("div",null,a.default.createElement("div",{className:"gallery"},a.default.createElement("img",{style:{width:"45%",margin:"0px 1%"},src:o.default.image.public.meetup_self}),a.default.createElement("img",{style:{width:"45%",margin:"0px 1%"},src:o.default.image.public.meetup_presentation})))),f=a.default.createElement("div",null,a.default.createElement("div",{className:"illustration"},a.default.createElement("iframe",{style:{height:"500px",width:"100%"},src:"https://www.youtube.com/embed/1wib7JQEZ78"})),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"About TRANS"),a.default.createElement("div",null,"TRANS Conference is a platform where entrepreneurs, experts, investors, and students can exchange ideas, which was created by H. Spectrum, a startup incubator/accelerator focusing on the healthcare sector. As we had a chance to join in this forum, we decided to present our project to people in the healthcare field as many as possible, hoping to get some feedback from them and improve our project."),a.default.createElement("div",null,a.default.createElement("div",{className:"gallery"},a.default.createElement("img",{className:"illust-image",src:o.default.image.public.trans})))),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Discussion and feedback"),a.default.createElement("div",null,"During TRANS, we found that people who are interested in our project are from multiple different fields and not just professionals. We’ve met students, bacterial professionals, fishery companies and some previous iGEM teams in Taiwan, whose backgrounds are medical, biology, microbiology, culture studies, medical engineering, and electrical engineering."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.public.discuss_1})),a.default.createElement("div",{className:"part"},a.default.createElement("div",null,"After the discussion, we sorted out the most common questions from our listeners as below:"),a.default.createElement("div",null,a.default.createElement("div",null,"1. What is the principle of MV-kit and what’s its difference between PCR analysis?"),a.default.createElement("div",null,"2. Biosafety such as being eaten by people and genetic mutation of E.coli."),a.default.createElement("div",null,"3. Will engineered genes exchange with bacterial organisms in the environment, causing genetic recombination problems because of the complex environment?"),a.default.createElement("div",null,"4. E.coli residual problems."),a.default.createElement("div",null,"5. Is engineered E.coli regulation acceptable? GMO products in Taiwan are still very opposed, how to persuade?"),a.default.createElement("div",null,"6. The growth and decline effect of E. coli will still have resistance problems in the future. Is there any solution?"))),a.default.createElement("div",{className:"part"},a.default.createElement("div",null,"And we also got some feedback and advice:"),a.default.createElement("div",null,a.default.createElement("div",null,"1. The mechanism of STAR/anti-STAR can also be applied to the sterilization of intensive care units."),a.default.createElement("div",null,"2. We could transform our target to a smaller range to simplify our problem, such as moving our environment from outdoor to indoor."),a.default.createElement("div",null,"3. In the future, if our growth and decline mechanism success, we could experiment our genes by intelligent shrimp culturing."),a.default.createElement("div",null,"4. To solve the problem of drug resistance, try adding multiple attack lines to E. coli."),a.default.createElement("div",null,"5. A company called ID Water which major in water quality testing suggested us to cooperate with them and tried to start an undertaking."))),a.default.createElement("div",{className:"gallery"},a.default.createElement("img",{style:{width:"30%",margin:"0px 1%"},src:o.default.image.public.discuss_2_1}),a.default.createElement("img",{style:{width:"30%",margin:"0px 1%"},src:o.default.image.public.discuss_2_2}),a.default.createElement("img",{style:{width:"30%",margin:"0px 1%"},src:o.default.image.public.discuss_2_3})),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"innerpart"},"After digesting the advice, we decided to modify our project in detail by meeting some of the professional companies, which we have saved the contacting information. First, we emailed to ID Water, asking for a chance to visit their company, hoping to have a chance to communicate some detecting methods. For example, we have trouble in analyzing with fluorescence in a bioreactor, which is part of their technique in water analyzing. ID Water thus told us that their cooperator, IGC-BIO, a healthcare company in Taiwan, is also researching in the method of interaction of bacteria, so we also plan to visit them someday."),a.default.createElement("div",{className:"innerpart"},"We could also meet Hin-Chung Wong professor in Soochow University, who is a profession in Vibrio field, for more detailed information about Vibrio to perfect our project. And we have checked again that our project doesn’t violate the iGEM rule. In the future, if we want to test our bacteria, LiveStrong Biomedical Technology promised us that they would provide a shrimp culturing sink for us to practice our experiment. Finally, thank for those who gave us lots of precious advice."),a.default.createElement("div",{className:"gallery"},a.default.createElement("img",{style:{width:"45%",margin:"0px 1%"},src:o.default.image.public.discuss_3_1}),a.default.createElement("img",{style:{width:"45%",margin:"0px 1%"},src:o.default.image.public.discuss_3_2}))))),d=a.default.createElement("div",null,a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.public.xgem})),a.default.createElement("div",null,"Interacting with people from different backgrounds is amazing and we do cherish every moment we engaged with other iGEM teams to exchange ideas as well as share techniques. For us, bringing the people across the world together to brainstorm about synthetic biology and how to make this world a better place is the magic of iGEM. Therefore, we established a platform on Slack named XGEM. On XGEM, teams can share information and ask questions immediately. So far, 18 teams from 14 different countries have joined XGEM. More about the teams joining XGEM:"),a.default.createElement("div",null,a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.public.chart_1})),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.public.chart_2})),a.default.createElement("div",{className:"illustration",style:{width:"70%"}},a.default.createElement("img",{className:"illust-image",src:o.default.image.public.chart_3}))),a.default.createElement("div",null,"Its really exciting to have 19 teams from all over the world to interact on XGEM. Some teams have posted their questionnaires related to their project or shared what they had done. We really appreciate your participation and can’t wait to meet these teams in person at Giant Jamboree!")),p={viewbar:{Title:"Public",Subtitle:"Meetups, conferences, and online platform",image:o.default.image.topfull.public},content:{Introducation:"Due to the fact that the synthetic biology is a brand-new field for the general public. Consequently, creating an enlarging division of knowledge between the general public and synthetic biology is necessary, and thus the public engagement is crucial for bridging the gap. The following are the activities we attended this year. We learned so much from the public, our friends and other iGEM teams. 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/** @license React v16.4.2

* react-is.production.min.js
*
* Copyright (c) 2013-present, Facebook, Inc.
*
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Therefore, we aim to create an IoT detecting device which can collect the water sample automatically and periodically. Then it is able to detect the concentration of specific bacteria with high specificity. For instance, it can detect the concentration of 100 CFU/mL of ",a.default.createElement("span",{className:"special-name"},"Vibrio")," within 24 hours[4,5], and informs fisherman regularly. It has the potential to reduce the financial loss of our aquaculture industry. ")),f=a.default.createElement("div",null,a.default.createElement("div",{className:"reference"},"1. Ching-Ying Deng, Meei-Mei Chen, Ju-Hsien Peng, Yu-Chiao Yang, Chih-Chien Chang, Hui-Ping Tsai 2014 The Investigation on the Infection of Francisellosis in Farmed Tilapia in Taoyuan and Hsinchu Areas. Journal of Taiwan Fisheries Research,22 (1): 81-91"),a.default.createElement("div",{className:"reference"},"2. https://news.un.org/en/audio/2014/05/588932, UN NEWS. 19 May 2014 "),a.default.createElement("div",{className:"reference"},"3. H-J Ju, M-Y Huang, H-C Liu, L-W Tseng, C-L Pan and C-I Chang, 2016 Screening Bacteria Antagonistic toward Aquatic Pathogenic Vibrio spp. from Marine Fishes and Shrimp. Journal of Taiwan Fisheries Research, 24 (1): 37-50"),a.default.createElement("div",{className:"reference"},"4. Chin-I Chan, et al., 2011, https://www.tfrin.gov.tw/dl.asp?fileName=17111695971.pdf"),a.default.createElement("div",{className:"reference"},"5. http://www.tfrin.gov.tw/friweb/frienews/enews0118/h1.html")),d={viewbar:{Title:"Description",Subtitle:"A solution to the problem of the abuse of antibiotics in aquaculture",image:o.default.image.topfull.description},content:{Introduction:"",parts:[{Title:"Motivation",Description:u},{Title:"Solution",Description:c},{Title:"Reference",Description:f}]}},p=function(e){function t(){return function(e,t){if(!(e instanceof t))throw new TypeError("Cannot call a class as a function")}(this,t),function(e,t){if(!e)throw new ReferenceError("this hasn't been initialised - super() hasn't been called");return!t||"object"!=typeof t&&"function"!=typeof t?e:t}(this,(t.__proto__||Object.getPrototypeOf(t)).apply(this,arguments))}return function(e,t){if("function"!=typeof t&&null!==t)throw new TypeError("Super expression must either be null or a function, not "+typeof t);e.prototype=Object.create(t&&t.prototype,{constructor:{value:e,enumerable:!1,writable:!0,configurable:!0}}),t&&(Object.setPrototypeOf?Object.setPrototypeOf(e,t):e.__proto__=t)}(t,i.Component),r(t,[{key:"render",value:function(){return a.default.createElement(l.default,d)}}]),t}();t.default=p},function(e,t,n){"use strict";Object.defineProperty(t,"__esModule",{value:!0}),t.default=void 0;var r=function(){function e(e,t){for(var n=0;n<t.length;n++){var r=t[n];r.enumerable=r.enumerable||!1,r.configurable=!0,"value"in r&&(r.writable=!0),Object.defineProperty(e,r.key,r)}}return function(t,n,r){return n&&e(t.prototype,n),r&&e(t,r),t}}(),i=n(0),a=s(i),o=s(n(4)),l=s(n(8));function s(e){return e&&e.__esModule?e:{default:e}}var u=a.default.createElement("div",null,a.default.createElement("div",null,"Our project this year is to design an engineered E.coli to co-culture with pathogenic Vibrio and create a prey-predator system, also called bio-control, to control Vibrio population in order to create an equilibrium between Vibrio and engineered E.coli. Therefore Vibrio population can be controlled in a particular concentration which will not able to infect marine organism such as shrimp, fish…etc."),a.default.createElement("div",null,a.default.createElement("div",{className:"illustration"},a.default.createElement("div",{className:"illust-image-wrapper"},a.default.createElement("img",{className:"illust-image",src:o.default.image.design.exp})),a.default.createElement("div",{className:"undertext"},"Fig 1. exponential growth of vibrio without constraint in fish farm")),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.design.equlibrium}),a.default.createElement("div",{className:"undertext"},"Fig 2. Vibrio population controlled by engineered E.coli forming a equilibrium")),a.default.createElement("div",null)),a.default.createElement("div",{className:"illustration"},a.default.createElement("div",{className:"illust-image-wrapper"},a.default.createElement("img",{className:"illust-image",src:o.default.image.design.shrimp})),a.default.createElement("div",{className:"undertext"},"Fig 3. Infected shrimp")),a.default.createElement("div",null,"Hence the goal of wetlab this year is going to design a gene circuit which can make engineered E.coli to secrete anti-vibrio substances[1] while vibrio concentration exceed hazard concentration, and engineered E.coli suicide[2] while vibrio concentration is low to avoid bio-safety concern."),a.default.createElement("div",{className:"illustration"},a.default.createElement("div",{style:{margin:"auto",width:"500px"}},a.default.createElement("img",{className:"illust-image",src:o.default.image.design.cycle}))),a.default.createElement("div",{className:"part"}," Our gene design can be divided into 3 parts. First is the “detection” part makes our E.coli sense the population of vibrio and respond to it.  Second, the “kill switch” part compares the level of 2 bacteria and switches our E.coli  between modes of secreting anti-vibrio substances and suicide substance. “Growth inhibitor” part includes a toxic protein (so called Colicin M) and a toxic small RNA (tsRNA)."),a.default.createElement("div",{className:"part"}," Though we want to conduct Vibrio related experiments in our lab, our lab belongs to BSL1 level and we can’t cultivate Vibrio. Therefore, we use 2 different kinds of E. coli. to simulate our succession model with one being as fake Vibrio, and another being as engineered E. coli. ")),c=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},"ACE is an abbreviation of “Automatic Color-detecting Examiner” which is able to provide a stable environment for any color-changing-related test. All of the processes are executed and the information of each color will be digitized automatically. Finally, we create our own database to store the data and analyze them more precisely. Combining with IoT technology, all of our data will be recorded and analyzed in our cloud database and be returned to our clients, which allows them to take control of the results."),a.default.createElement("div",{className:"part"},"In order to prove our ACE can be used in reality to solve the Vibrio problem in our society, we make a collaboration with the “Fisheries Research Institute” of the Taiwan government. 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OD 600 Reference point"),a.default.createElement("div",null,"LUDOX CL-X (45% colloidal silica suspension) is used as a single point reference to obtain a conversion factor to transform absorbance (Abs600) data from our plate reader into a comparable OD600 measurement as would be obtained in a spectrophotometer. We will get the data for OD600 of the H2O and LUDOX. The corrected Abs600 is calculated by subtracting the H2O reading. To convert measured Abs600 to OD600 is to let Reference OD600 divided by Abs600.")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"2. Particle Standard Curve"),a.default.createElement("div",null,"We prepared a dilution series of monodisperse silica microspheres and measured the Abs600 in a plate reader. The size and optical characteristics of these microspheres are similar to cells, and there was a known amount of particles per volume. This measurement allows us to construct a standard curve of particle concentration which can be used to convert Abs600 measurements to an estimated number of cells.")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"3. Fluorescence standard curve"),a.default.createElement("div",null,"Absolute fluorescence values cannot be directly compared from one instrument to another. In order to compare the fluorescence output of test devices between teams, it was necessary to create a standard fluorescence curve.")))),f=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},a.default.createElement("div",null,"First, we transformed Escherichia coli DH5α with these following plasmids."),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",null,"Negative control   BBa_R0040    Kit Plate 7 Well 2D"),a.default.createElement("div",null,"Positive control   BBa_I20270   Kit Plate 7 Well 2B"),a.default.createElement("div",null,"Test Device 1   BBa_J364000  Kit Plate 7 Well 2F"),a.default.createElement("div",null,"Test Device 2   BBa_J364001  Kit Plate 7 Well 2H"),a.default.createElement("div",null,"Test Device 3   BBa_J364002  Kit Plate 7 Well 2J"),a.default.createElement("div",null,"Test Device 4   BBa_J364007  Kit Plate 7 Well 2L"),a.default.createElement("div",null,"Test Device 5      BBa_J364008  Kit Plate 7 Well 2N"),a.default.createElement("div",null,"Test Device 6      BBa_J364009  Kit Plate 7 Well 2P"))),a.default.createElement("div",{className:"part"},"Secondly, two colonies from each plate were picked and inoculated in LB medium containing chloramphenicol overnight for 16-18 hours at 37°C and 220 rpm."),a.default.createElement("div",{className:"part"},"Finally, we measured the OD600 and fluorescence of transformed cells according to the protocol after 0, 6 hours. 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Outside of iGEM, he is now joining in an immunology laboratory and doing some researches about immune checkpoints, PD-1. Apart from the university studies, he is also active in diverse activities. For example, he has participated in several Chinese calligraphy contests in Taiwan and won many prizes.",contact:{facebook:"https://www.facebook.com/tysh0106",linkedin:"https://www.linkedin.com/in/tysh0106/"}},{name:"Kevin Wang",image:""+C.default.image.member.Kevin_Wang,position:"Wetlab",department:"Materials Science and Engineering/ Physics",description:"I am Kevin Wang, a sophomore student study in Material Sciences and Physics. \n      During iGEM, I design the gene circuit and most of gene fragments of our project this year, and including trouble shooting. This summer, I spent lots of time doing digestion-ligation, transform and PCR. Although failure make me feel frustration, I never give up till success. 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They kindly provided us with their “Multiple-Vibrio Detecting Kit, MV-kit” and we integrated their MV-kit with our system to develop a brand-new",o.default.createElement("a",{href:"https://2018.igem.org/Team:NTHU_Taiwan/Application"}," Automatic Vibrio-detecting System"),".")),o.default.createElement("div",{className:"part"},"Through the optimization of ACE, we accelerate the detection rate of MV-kit. With our ACE, it becomes 2 times faster than the original MV-kit and the minimal detectable Vibrio concentration is about 100 CFU/ mL within 24 hours. In summary, we combine the traditional detecting methods with the cutting-edge technology to achieve a better effect. In addition, following with our IoT system makes the water quality testing system in aquatic agriculture industry step forward to the whole new world of “intelligent aquaculture”."),o.default.createElement("div",{className:"illustration"},o.default.createElement("img",{className:"illust-image",src:s.default.image.drylab_overview.plate}),o.default.createElement("div",{className:"undertext"},"The color gradient changing process"))),m=o.default.createElement("div",null,o.default.createElement("div",null,"Components list and cost on ",o.default.createElement("a",{href:" https://docs.google.com/spreadsheets/d/1NsoRao-FtywW1KCeFYrxMIjJG3kVGQRcg_fIhxjn7ZQ/edit?usp=sharing"},"Google drive")),o.default.createElement("div",null,"3D printing files on ",o.default.createElement("a",{href:"https://drive.google.com/drive/folders/1tPQEVtNHB_hzhalPlwZu4bzyQVSDUPkN?usp=sharing"},"Google Drive"))),g={viewbar:{Title:"Drylab",Subtitle:"ACE- Automatic Color-detecting Examiner",image:s.default.image.topfull.drylab_overview,Underimage:f},content:{Introduction:d,parts:[{Title:"ACE",Description:p},{Title:"Application",Description:h},{Title:"File download",Description:m}]}},v=(0,l.inject)("appState")(r=(0,l.observer)(r=function(e){function t(e){return function(e,t){if(!(e instanceof t))throw new TypeError("Cannot call a class as a function")}(this,t),function(e,t){if(!e)throw new ReferenceError("this hasn't been initialised - super() hasn't been called");return!t||"object"!=typeof t&&"function"!=typeof t?e:t}(this,(t.__proto__||Object.getPrototypeOf(t)).call(this,e))}return function(e,t){if("function"!=typeof t&&null!==t)throw new TypeError("Super expression must either be null or a function, not "+typeof t);e.prototype=Object.create(t&&t.prototype,{constructor:{value:e,enumerable:!1,writable:!0,configurable:!0}}),t&&(Object.setPrototypeOf?Object.setPrototypeOf(e,t):e.__proto__=t)}(t,a.Component),i(t,[{key:"render",value:function(){return o.default.createElement(u.default,g)}}]),t}())||r)||r;t.default=v},function(e,t,n){"use strict";Object.defineProperty(t,"__esModule",{value:!0});var r=function(){function e(e,t){for(var n=0;n<t.length;n++){var r=t[n];r.enumerable=r.enumerable||!1,r.configurable=!0,"value"in r&&(r.writable=!0),Object.defineProperty(e,r.key,r)}}return function(t,n,r){return n&&e(t.prototype,n),r&&e(t,r),t}}(),i=n(0),a=s(i),o=(s(n(10)),s(n(4))),l=s(n(8));function s(e){return e&&e.__esModule?e:{default:e}}var u=a.default.createElement("div",null,a.default.createElement("div",null,"The reaction platform mainly consists of three parts: "),a.default.createElement("div",{className:"innerpart"},"1. The 96-Well Microtitration Plate for sample storage and reaction "),a.default.createElement("div",{className:"innerpart"},"2. Light blocking base to black out the area we’re not using to make it easier for camera to detect the color changes of each well"),a.default.createElement("div",{className:"innerpart"},"3. Auto moving lid to prevent the reactive reagent from evaporating")),c=a.default.createElement("div",null,a.default.createElement("div",null,"After extracting a sample from the fish farm, the sample will be dripped into the 96 -well plate. In the plate, we put in the mixture of MV-kit buffer for the sample to react. There are several reasons we chose to use 96-well plates instead of eppendorf or 6/24/48 well plate."),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"innerpart"},"1. Eppendorf has an unremovable lid, which is not convenient for our auto sampling design."),a.default.createElement("div",{className:"innerpart"},"2. The base of eppendorf is sharp and liquid will not distribute evenly, which is not suitable for color sensing while 96-well plates are flat."),a.default.createElement("div",{className:"innerpart"},"3. The volume of the 96-well plate is small enough for us to put the sample in, while the other plates are too big."),a.default.createElement("div",{className:"innerpart"},"4. It can contain the most samples in one single plate. In that way, we can reduce the rate of changing new plates.")),a.default.createElement("div",{className:"illustration"},a.default.createElement("table",{className:"table"},a.default.createElement("tbody",null,a.default.createElement("tr",null,a.default.createElement("th",{className:"tablehead"},"Number of well"),a.default.createElement("th",{className:"tablehead"},"Radius (mm) "),a.default.createElement("th",{className:"tablehead"},"Max Volume (mL) "),a.default.createElement("th",{className:"tablehead"},"Growing Area (cm2)")),a.default.createElement("tr",null,a.default.createElement("td",{className:"tabledata"},"96"),a.default.createElement("td",{className:"tabledata"},"4.5 "),a.default.createElement("td",{className:"tabledata"},"0.36"),a.default.createElement("td",{className:"tabledata"},"0.16")),a.default.createElement("tr",null,a.default.createElement("td",{className:"tabledata"},"48"),a.default.createElement("td",{className:"tabledata"},"10.2"),a.default.createElement("td",{className:"tabledata"},"1.7"),a.default.createElement("td",{className:"tabledata"},"0.8")),a.default.createElement("tr",null,a.default.createElement("td",{className:"tabledata"},"24"),a.default.createElement("td",{className:"tabledata"},"15.6"),a.default.createElement("td",{className:"tabledata"},"3.4"),a.default.createElement("td",{className:"tabledata"},"1.9")),a.default.createElement("tr",null,a.default.createElement("td",{className:"tabledata"},"12"),a.default.createElement("td",{className:"tabledata"},"22.1"),a.default.createElement("td",{className:"tabledata"},"6.9"),a.default.createElement("td",{className:"tabledata"},"3.8")),a.default.createElement("tr",null,a.default.createElement("td",{className:"tabledata"},"6"),a.default.createElement("td",{className:"tabledata"},"34.8"),a.default.createElement("td",{className:"tabledata"},"16.8"),a.default.createElement("td",{className:"tabledata"},"9.5")))),a.default.createElement("div",{className:"undertext"},"reference: http://www.dxy.cn/bbs/topic/9584997"),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.platform.transparent})),a.default.createElement("div",{className:"undertext"},"reference: http://www.carelifetech.com.tw/?96%E5%AD%94%E7%9B%A4-96-well-plate,127"))),f=a.default.createElement("div",null,a.default.createElement("div",null,"Since we need to use the camera to detect the color changes of each well, we need to create an environment that can make the color as clear as possible.  Therefore, we use 3D printing and polylactic acid (PLA) to print out a platform that blacks out all the area except for the 24 holes of each plate that have our sample in."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.platform.block}))),d=a.default.createElement("div",null,a.default.createElement("div",null,"Since the MV-kit buffer and solution might evaporate, we need to construct an auto moving lid. The lid has 48 holes on it, each hole is 9.1 mm next to the well. This lid is connected with a gear and a stepper motor. Every time the sample is being dripped into the well, the motor will move the lid by 9.1 mm. The gear system and the base of the motor are also made by 3D printing. Moreover, we’ve designed the lid to be detachable so that the farmer can remove the lid when they are going to substitute the 96 well plates with a new one every two weeks. "),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.platform.moving}))),p={viewbar:{Title:"Platform",Subtitle:"Plate and the moving lid",image:o.default.image.topfull.platform},content:{Introduction:u,parts:[{Title:"96-Well Microtitration",Description:c},{Title:"Light blocking",Description:f},{Title:"Moving lid",Description:d}]}},h=function(e){function t(){return function(e,t){if(!(e instanceof t))throw new TypeError("Cannot call a class as a function")}(this,t),function(e,t){if(!e)throw new ReferenceError("this hasn't been initialised - super() hasn't been called");return!t||"object"!=typeof t&&"function"!=typeof t?e:t}(this,(t.__proto__||Object.getPrototypeOf(t)).apply(this,arguments))}return function(e,t){if("function"!=typeof t&&null!==t)throw new TypeError("Super expression must either be null or a function, not "+typeof t);e.prototype=Object.create(t&&t.prototype,{constructor:{value:e,enumerable:!1,writable:!0,configurable:!0}}),t&&(Object.setPrototypeOf?Object.setPrototypeOf(e,t):e.__proto__=t)}(t,i.Component),r(t,[{key:"render",value:function(){return a.default.createElement(l.default,p)}}]),t}();t.default=h},function(e,t,n){"use strict";Object.defineProperty(t,"__esModule",{value:!0});var r=function(){function e(e,t){for(var n=0;n<t.length;n++){var r=t[n];r.enumerable=r.enumerable||!1,r.configurable=!0,"value"in r&&(r.writable=!0),Object.defineProperty(e,r.key,r)}}return function(t,n,r){return n&&e(t.prototype,n),r&&e(t,r),t}}(),i=n(0),a=s(i),o=(s(n(10)),s(n(8))),l=s(n(4));function s(e){return e&&e.__esModule?e:{default:e}}var u=a.default.createElement("div",null,"For the results of color-changing, we write a program to detect them. In our device, there are 24 samples processing the chemical reaction simultaneously. During the reaction, we want to get all samples’ color values immediately, so we adopt image detection executed by Raspberry Pi to analyze the whole status directly. The following are three parts for our image detection, which is how to do, advantages, and problems respectively."),c=a.default.createElement("div",null,a.default.createElement("div",null,"The following is the materials image detection needs"),a.default.createElement("table",{className:"table"},a.default.createElement("tbody",null,a.default.createElement("tr",null,a.default.createElement("td",{className:"tabledata"},"Raspberry Pi 3 Model B v1.3"),a.default.createElement("td",{className:"tabledata"},"1")),a.default.createElement("tr",null,a.default.createElement("td",{className:"tabledata"},"Webcam Logitech C270"),a.default.createElement("td",{className:"tabledata"},"1")),a.default.createElement("tr",null,a.default.createElement("td",{className:"tabledata"},"Wide-angle lens 37mm 0.6X"),a.default.createElement("td",{className:"tabledata"},"1")))),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:l.default.image.imaging.materials})),a.default.createElement("div",{className:"part"},"The core of image detection is executed by Raspberry Pi and the program language we use is python. With a camera and wide-angle lens, it can capture the scene in a relatively lower height. To do the image processing, OpenCV, open source library, is the best choice. Because the program will send warning message to the users, for simplicity and stability, we choose MQTT to be the transport protocol."),a.default.createElement("div",{className:"part"},"The following three steps are the processes our image detection works on. First, it captures the scene using a camera in a specified time. Second, the image is analyzed immediately after one is captured and the results are record. Third, after the analysis, if there is a color-changing sample, it will send the warning message to the users. All the steps are operated recursively until all the samples are analyzed.")),f=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},"For our image detection, we have three advantages, which are accuracy, automation, and flexibility, respectively. With these three, it can match our goal of designing ACE, i.e., automation and also increase usability compared to other devices used in the laboratory nowadays."),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"1. Accuracy"),a.default.createElement("div",{className:"innerpart"},"Because our 24 samples are close to each other, the accuracy is necessary to avoid the wrong result of analysis. With image detection, we can set the section where we want to analyze corresponding to the samples on the picture. Therefore, if all samples are located in advance carefully, we can get high accuracy for the results.")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"2. Automation"),a.default.createElement("div",{className:"innerpart"},"Automation is our goal of designing the device. To achieve this, the program can automatically do the following things, i.e., capturing the scene, analyzing the captured image, recording the results, and sending the warning messages to users. With the four steps as mentioned above operated cyclically, the program achieves automation.")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"3. Flexibility"),a.default.createElement("div",{className:"innerpart"},"The section program analyzes on the captured image can be modified whenever the captured one is changed. Therefore, we don’t have to fix the position of all samples. Due to this advantage, any type of sample can be analyzed accurately."))),d=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},"During the development of image detection, we had made some choices to solve many problems, including the specific practice of detection, development board, the algorithm to analyze the images, and the environment light. The following describes each problem, and the reasons why we made these choices."),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"1. Image detection vs. Color sensor"),a.default.createElement("div",{className:"innerpart"},"Initially, color sensor is our direction to design the device. Moving the color sensor to the top of each sample can get the color of that. However, our samples are close to each other, and the resolution of a color sensor is not high enough to detect the color of single sample accurately, so the direction is changed to image detection with the camera capturing the scene. In this way, it analyzes the image directly, the accuracy increases by a large margin.")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"2. Raspberry Pi vs. Other development boards"),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:l.default.image.imaging.board})),"For the development boards, there are three choices, Arduino, Udoo Neo, and Raspberry Pi. Considering the efficiency, Arduino might not be able to run OpenCV effectively. The another consideration is the cost. Because we want to design a low-cost device cheaper than the similar machine sold on the market, the price of Udoo Neo is too expensive for us. Therefore, we chose Raspberry Pi to be our execution core.")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"3. Simplified location vs. Houghcircles"),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:l.default.image.imaging.algorithm})),"In the beginning, we planned to use OpenCV built-in function Houghcircles to do image detection. The detail of the function is that it can find circles on the image automatically. It completely meets our expectation. However, the distance and angle of each sample from the camera is different. As a result, the shape of the sample is not a circle on the captured image and leads to wrong detection by Houghcircles. For this reason, we changed the method and decided to locate the samples manually to get a high accuracy.")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"4. LED strip vs. Big bulb"),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:l.default.image.imaging.light})),"In order to prevent the influence of environment outside the device, ACE is designed to be enclosed. Therefore, it is completely dark inside the device and needs a light source to do image detection. In the beginning, we thought a big bulb is simple and suitable. Nevertheless, after we tested, we found the light on the image is not even, it would result in the wrong analysis. As a result, we chose LED strip. Its light is not only even but also bright enough.")),p=a.default.createElement("div",null,"You can download the source of our program on",a.default.createElement("a",{href:"https://github.com/igem-tw-nthu/opencv"}," Github")),h={viewbar:{Title:"Imaging",Subtitle:"Imaging by Raspberry Pi with OpenCV",image:l.default.image.topfull.imaging},content:{Introduction:u,parts:[{Title:"How to Do",Description:c},{Title:"Advantages",Description:f},{Title:"Problems",Description:d},{Title:"Code Download",Description:p}]}},m=function(e){function t(){return function(e,t){if(!(e instanceof t))throw new TypeError("Cannot call a class as a function")}(this,t),function(e,t){if(!e)throw new ReferenceError("this hasn't been initialised - super() hasn't been called");return!t||"object"!=typeof t&&"function"!=typeof t?e:t}(this,(t.__proto__||Object.getPrototypeOf(t)).apply(this,arguments))}return function(e,t){if("function"!=typeof t&&null!==t)throw new TypeError("Super expression must either be null or a function, not "+typeof t);e.prototype=Object.create(t&&t.prototype,{constructor:{value:e,enumerable:!1,writable:!0,configurable:!0}}),t&&(Object.setPrototypeOf?Object.setPrototypeOf(e,t):e.__proto__=t)}(t,i.Component),r(t,[{key:"render",value:function(){return a.default.createElement(o.default,h)}}]),t}();t.default=m},function(e,t,n){"use strict";Object.defineProperty(t,"__esModule",{value:!0});var r=function(){function e(e,t){for(var n=0;n<t.length;n++){var r=t[n];r.enumerable=r.enumerable||!1,r.configurable=!0,"value"in r&&(r.writable=!0),Object.defineProperty(e,r.key,r)}}return function(t,n,r){return n&&e(t.prototype,n),r&&e(t,r),t}}(),i=n(0),a=u(i),o=(u(n(10)),u(n(4))),l=u(n(8)),s=u(n(136));function u(e){return e&&e.__esModule?e:{default:e}}var c=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},a.default.createElement("div",null,"For most of the detection reagent, they can only react with the small amount of sample each time. Taking MV-kit (Multiple Vibrio Detecting Kit) that we’re using for vibrio detection for example, it needs to have fix amount of 35μl sample fluid being collected every time so that we can get the better result of the later on reaction with the reagent. We choose mini peristaltic pump to fulfill our need. There are a few advantages of choosing peristaltic pump as our dispensing media[1]:"),a.default.createElement("div",null,a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",{className:"part-title"},"1. Accurate metering "),a.default.createElement("div",null,"Peristaltic pumps use flexible tubing to run through rollers in the pump head. As fluid goes through the pump head, packet of fluids can be formed. Simply by determining the size of the tubing and the speed of the roller turning in the pump head, the accurate metering of the media can be achieved.")),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",{className:"part-title"},"2. Self priming "),a.default.createElement("div",null,"Peristaltic pumps can draw fluid into the tubing when starting dry while other pumps require user to fill pump and suction line with fluid before use which can be inconvenient and also create potential for contamination.")),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",{className:"part-title"},"3. Non-siphoning "),a.default.createElement("div",null,"A peristaltic pump is non-siphoning which means that it prevents back flow into the system when the pump stop turning. This promotes accuracy during dispensing.")))),a.default.createElement("div",{className:"part"},a.default.createElement("div",null,"As a result, peristaltic pumps are often used for applications involving metering, dispensing and precise general transfer. Here, we use it in our device and we get the stl. files from the open source of the YouTube channels called “electronic crafts”[2]. The dimension of our peristaltic pump is 31 x 31 x 11 (mm), and the detail of other components are showed in Table1 and Figure 1. We printed the main components out by using the SLA 3D printer."),a.default.createElement("table",{className:"table"},a.default.createElement("tbody",null,a.default.createElement("tr",null,a.default.createElement("th",{colSpan:"6",className:"tablehead"},"Peristaltic Pump")),a.default.createElement("tr",null,a.default.createElement("td",{className:"tabledata"},"Dimension"),a.default.createElement("td",{className:"tabledata"},"Tube diameter"),a.default.createElement("td",{className:"tabledata"},"Tube material"),a.default.createElement("td",{className:"tabledata"},"Number of rollers"),a.default.createElement("td",{className:"tabledata",style:{margin:"0px 10px"}},"Bearing"),a.default.createElement("td",{className:"tabledata"},"Motor")),a.default.createElement("tr",null,a.default.createElement("td",{className:"tabledata"},"31 x 31 x 11 (mm)"),a.default.createElement("td",{className:"tabledata"},a.default.createElement("div",null,"OD: 3mm"),a.default.createElement("div",null,"ID: 1.5mm")),a.default.createElement("td",{className:"tabledata"},"Silicone rubber"),a.default.createElement("td",{className:"tabledata"},"three"),a.default.createElement("td",{className:"tabledata"},a.default.createElement("div",null,"OD: 9mm"),a.default.createElement("div",null,"ID: 3mm")),a.default.createElement("td",{className:"tabledata"},a.default.createElement("div",null,"28BYJ-48"),a.default.createElement("div",null,"12V"))))),a.default.createElement("div",{className:"undertext"},"Table 1"),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.sampling.pump}),a.default.createElement("div",{className:"undertext"},"Figure 1"))),a.default.createElement("div",{className:"part"},"Because peristaltic pumps deliver fixed amounts of fluid with each pass of a roller over the tube, the size of the tube has a direct effect on the amount of fluid delivered. Well-designed pumps are engineered to work with an optimum tubing size or range of tubing sizes, which take into account both the tube's inside diameter and wall thickness. The inside diameter determines the amount of fluid delivered with each turn of the rotor, while the wall thickness affects the tubing's ability to spring back to its original shape after each compression, which has a great influence on the overall life of the tubing[3]."),a.default.createElement("div",{className:"part"},"We choose the tube with inner diameter 2mm and wall thickness 1mm. Three bearings are used as the rollers in our peristaltic pump. The outer diameter of the bearing is 9mm and the inside diameter is 3mm. The peristatic pump after assembling can be seen in Figure 2. We can also calculated the theoretical discharge to know the theoretical performance of the peristaltic pump by using the equation below[4],"),a.default.createElement("div",{className:"part",style:{display:"flex",alignItems:"center"}},a.default.createElement("div",null,a.default.createElement("div",null,a.default.createElement(s.default.Provider,null,a.default.createElement(s.default.Node,{inline:!0,formula:"flowrate (μl/s) = V \\cdot L \\cdot N \\cdot RPM \\cdot10^3 \\cdot 60"}))),a.default.createElement("div",null,"V= Volume of occluded tubing (mm^3/mm)"),a.default.createElement("div",null,"L= Tubing length that will be occluded by pump rollers (mm)"),a.default.createElement("div",null,"N= Number of rollers on the rotor."),a.default.createElement("div",null,"RPM= pump rpm")),a.default.createElement("div",{style:{marginLeft:"auto",padding:"0 20px"}},a.default.createElement("img",{style:{height:"180px"},src:o.default.image.sampling.full_pump}),a.default.createElement("div",{className:"undertext"},"Figure 2"))),a.default.createElement("div",{className:"part"},a.default.createElement("div",null,"We have done multiple testing and calculated the weight of each droplet by measuring the weight of every ten droplets to get the average value. The average weight of each droplet is 0.035+-0.00072μl, and the rest of the measuring records are showed in Figure 3. This is exactly what we need for sample preparing for the MV-kit in our experiments."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.sampling.list}),a.default.createElement("div",{className:"undertext"},"Figure 3"))),a.default.createElement("div",{className:"part",style:{display:"flex",alignItems:"center"}},a.default.createElement("div",{style:{marginRight:"auto",padding:"0px 30px"}},a.default.createElement("img",{style:{height:"180px"},src:o.default.image.sampling.head}),a.default.createElement("div",{className:"undertext"},"Figure 4")),a.default.createElement("div",null,"Although we can almost get the precise amount of sample each time, it’s still hard to control the later droplets from dropping down cause of the gravity. Hence, we combine the pipette tip (Figure 4) at the end of the silicon tube to reduce the diameter and slow down the flowing rate. In addition, we also design the special lid with slopes in between to cover on the 96-well plate. We can utilize the slope design not only to drop the first few droplets that might be contaminated by the previous sample but also use them as the tunnels to collect the waste sample and remove all of them after the whole examination is done.")),a.default.createElement("div",{className:"illustration"},a.default.createElement("video",{className:"illust-image",autoPlay:!0,muted:!0,loop:!0},a.default.createElement("source",{src:o.default.video.lid,type:"video/mp4"})))),f=a.default.createElement("div",null,a.default.createElement("div",null,"We use two 12V 28BYJ-48 stepper motors to control the XY axis to move our sample to the specific coordinate and connect them to Arduino as well as uln2003 motor drivers to control the speed and time. The step angle of the motor is 5.625 degrees and the reduction ratio is 1/64, therefore, for one revolution it needs 4096 steps. The diameter of the gear mounted on the motor is 36mm, so for each step, it can linearly move forward or backward 1.76mm. The length between two wells on 96-well plate is about 18mm, thus, this motor can absolutely satisfy our need for our low-cost XY plane setting. "),a.default.createElement("div",{className:"illustration"},a.default.createElement("video",{className:"illust-image",autoPlay:!0,muted:!0,loop:!0},a.default.createElement("source",{src:o.default.video.sampling,type:"video/mp4"})))),d=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},"At first, we’re trying to set up the XY axis by using the DC motor to control it in considering of the low-cost budget. However, we found out that since the DC motor is categorized as analog motor, we can only drive them by sending power signals (two wire) to the motor to drive them. This would be tricky for us to do the sophisticated planned moves because we won’t know exactly where they may be located except trying to predict the moves over time within a relative margin of error(3-8%)[5]. Moreover, the friction between the gear rack and the motor make it even hard to control and need to build up more complex moves manually. "),a.default.createElement("div",{className:"part"},"On the other hand, stepper motors are digital motors, we can actually send position signals (four wire) to the motor to drive them. It’s possible for us to know exactly where they are at all times, and would be easier for us to do ‘repeat’ motion passes and have it return to a ‘home and start’ position precisely. For this reason even most sophisticated of a moves can be achieved. "),a.default.createElement("div",{className:"part"},"Due to all the reasons above, instead of making motion decision ‘on the fly’ at the rig, we then choose stepper motor as our driving motor.")),p=a.default.createElement("div",null,a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.sampling.result}))),h=a.default.createElement("div",null,a.default.createElement("div",{className:"reference"},"1. https://www.manufacturingchemist.com/news/article_page/Peristaltic_pumps__advantages_and_applications/74693"),a.default.createElement("div",{className:"reference"},"2. Peristaltic pump design: https://www.youtube.com/watch?v=J2KLq7485kM"),a.default.createElement("div",{className:"reference"},"3. https://archive-resources.coleparmer.com/TechInfo/HTMLFiles/PPTube_match.htm"),a.default.createElement("div",{className:"reference"},"4. Suhas R. Dhumal, Design and Development of Rotary Peristaltic Pump, International Journal of Science and Advanced Technology (ISSN 2221-8386)"),a.default.createElement("div",{className:"reference"},"5. https://support.dynamicperception.com/hc/en-us/articles/202986595-What-is-the-difference-between-DC-motors-and-Stepper-motors-")),m={viewbar:{Title:"Sampling",Subtitle:"Auto sampling with peristaltic pump ",image:o.default.image.topfull.sampling},content:{Introduction:"In our device, in order to let the following detected process work out properly, we first set up the automated sampling mechanism. 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However, we found that it is not bright enough, so we chose another kind of light."),a.default.createElement("div",{className:"part"},"After this, we choose the white light bar and connect it with Arduino. Since the voltage afforded by Arduino is 12V, it’s too bright to observe, so we apply the voltage-gain-module to get the appropriate voltage. Then we find out the white light may sometimes be difficult for us to tell apart whether the liquid changes color or not. After discussion, we change the white light to yellow light and the problem is solved. ")),c=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},'Our Vibrio detection device needs to be able to react in a stable 30 ° C environment. It is hoped that a method can be designed to heat up when the ambient temperature is high and cool down when the temperature is low. The cooling technologies used today are mainly the following: "high pressure gas expansion and cooling", "liquid evaporation and refrigeration", "gas vortex cooling", "semiconductor refrigeration" and "chemical cooling", and we choose "semiconductor cooling", which is a relatively easy to complete and within the capability range.'),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"First step"),a.default.createElement("div",{className:"innerpart"},"Common methods for cooling semiconductors include water cooling and fan cooling. Among them, due to our large architecture, water cooling is effective but too cumbersome and heavy. Limited by the space configuration, we use fan cooling. (Picture 4) By keep bringing the heat away, the temperature gradient of the chip become larger, and finally it will stop at a balanced point. We also add a fan on the cold side since we want the cold air to be uniformed inside the box. (Picture 5) Temperature-controlled circuits are more cumbersome and too complicated circuits are not suitable for use in our system, so we try to use temperature control module first. (Picture 6)"),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:l.default.image.environment.first}),a.default.createElement("div",{className:"undertext"})),a.default.createElement("div",{className:"innerpart"},"We found that as we put the cooling device in a cardboard box for testing, the result is not good since the space is not sealed and made the chip hard to cool down the environment."),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",{className:"innerpart"},"Needed materials:"),a.default.createElement("div",null,"two 5V-2wire fans, two small heat sinks, some thermal grease, one thermoelectric cooling chip provided by a national competition in Taiwan, temperature control module."))),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Second step"),a.default.createElement("div",{className:"innerpart"},"To enlarge the temperature difference, we add a more powerful fan on the hot side to make sure the rate of heat dissipation is enough for the rate of chip getting cold, which is a 12V-2wire one. As a result, the inbox environment could drop 3℃, but still didn’t reach our expectation."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:l.default.image.environment.second}),a.default.createElement("div",{className:"undertext"},a.default.createElement("div",null,"Figure 1: The air temperature inside the box drops from 26.6℃ to about 18℃ in 7 minutes, and maintains at 18℃ for 30 minutes. (cold device) The air temperature inside the box rises from 27.2℃ to 32.8℃ in about 6 minutes, and maintains at 32.8℃ for 15 minutes. (hot device)"),a.default.createElement("div",null,"Figure 2: When cold devices turn off, the air temperature inside the box rises from 27.2℃ to 32.8℃ in about 6 minutes, and maintains at 32.8℃ for 15 minutes. (cold device) When the hot device turn off, the air temperature inside the box drops from about 32.8℃ to 27.6℃ in 6 minutes, the same as the time to heat up the air. (hot device)"))),a.default.createElement("div",{className:"innerpart"},'During our experiment, we found that the temperature control module couldn’t provide a two-state system, which is hard to be designed in our project. Therefore, we try another way by using Arduino and relay module to control the temperature directly. Relay, also known as electric cymbal, is an electronic control device that has a control system (also known as an input circuit) and a controlled system (also known as an output circuit). It is usually used in automatic control circuits. It is actually used as an "automatic switch" that uses a small current to control large currents. Therefore, it plays the role of automatic adjustment, safety protection and conversion circuit in the circuit. Besides, we chose an IC called LM35 as our temperature sensor by transforming resistance into temperature. The wiring diagram is as below. (Picture 8)'),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",{className:"innerpart"},"Needed materials: "),"Cold device is combined by 12V-2wire fan, heat sink, thermal grease, thermoelectric cooling chip, thermal grease, heat sink, 5V-2wire fan (from up to down); Temperature controlling: Arduino for control, LM35 for sensing, relay module for switch.")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Final step"),a.default.createElement("div",{className:"innerpart"},"To reduce the unknown impact on the experiment, we use two cold device and one hot device to control the temperature. Hot device is similar to cold one, the only different is its two fans are all 5V-2wire, which is the device we used in the first period."),"  ",a.default.createElement("div",{className:"innerpart"},"This time we use two cold devices to find for the lowest temperature. Figure 1 shows the result we test cold or hot devices, and Figure 2 shows the changing after we turn off the devices."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:l.default.image.environment.final}),a.default.createElement("div",{className:"undertext"})),a.default.createElement("div",{className:"inner-part"},"After discussion, we decide to give styrofoam a try, we add it inside the box and do the rest of the experiment, includes add it or not add it. We finally decide to use two cold devices and one hot device without adding any material."),a.default.createElement("table",null),a.default.createElement("div",{className:"innerpart"},"We use Arduino connects with relay and temperature sensor to control the temperature. When the sensor detects the temperature lower than the setting, hot device is activated, it would heat up the air inside the device until it reaches the target temperature. Then, the hot device continues working that makes the temperature higher than the ideal one, this time, the cold devices turn on and cool down the environment. At the end, the temperature would be controlled in the range we have written in Arduino."),a.default.createElement("div",{className:"innerpart"},"In the beginning, we set the temperature to 27℃, which is a good condition for germs to grow. However, we neglect that there would be some the delay of temperature change between the air and the sensor, and the temperature around the sensor became higher than the environment, which affected the detecting correction. After we tidied up the wires, the problems disappeared and came into perfect."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:l.default.image.environment.end}),a.default.createElement("div",{className:"undertext"},"Figure 6: Temperature control simulation. The hot device turns on at the beginning, and the cold devices turn on when the temperature reaches 28.3℃, in about 6 minutes.")))),f=a.default.createElement("div",null,a.default.createElement("div",{className:"reference"},"1. Thermal Conductivity and Thermoelectric Power of Semiconductors. I. Terasaki, in Reference Module in Materials Science and Materials Engineering, 2016"),a.default.createElement("div",{className:"reference"},"2. Physics and Fundamental Theory. I. Terasaki, in Comprehensive Semiconductor Science and Technology, 2011"),a.default.createElement("div",{className:"reference"},"3. Thermoelectric cooling. JoséMeseguer, IsabelPérez-Grande, AngelSanz-Andrés, in Spacecraft Thermal Control, 2012"),a.default.createElement("div",{className:"reference"},"4. https://en.wikipedia.org/wiki/Thermoelectric_effect#/media/File:Thermoelectric_Cooler_Diagram.svg")),d={viewbar:{Title:"Environment",Subtitle:"Led Light, and peltier cooler",image:l.default.image.topfull.environment},content:{Introduction:"In order to make sure the observation results are stable and reliable, it’s necessary to control the experiment environment. Besides putting all the instruments inside our acrylic-made box (size: 20*30*35, unit: cm3), temperature and brightness are also important. In this case, we separate the “Environment” part into two sub-titles, temperature, and light. We will introduce more details in the following statements.",parts:[{Title:"Light",Description:u},{Title:"Temperature",Description:c},{Title:"Reference",Description:f}]}},p=function(e){function t(e){return function(e,t){if(!(e instanceof t))throw new TypeError("Cannot call a class as a function")}(this,t),function(e,t){if(!e)throw new ReferenceError("this hasn't been initialised - super() hasn't been called");return!t||"object"!=typeof t&&"function"!=typeof t?e:t}(this,(t.__proto__||Object.getPrototypeOf(t)).call(this,e))}return function(e,t){if("function"!=typeof t&&null!==t)throw new TypeError("Super expression must either be null or a function, not "+typeof t);e.prototype=Object.create(t&&t.prototype,{constructor:{value:e,enumerable:!1,writable:!0,configurable:!0}}),t&&(Object.setPrototypeOf?Object.setPrototypeOf(e,t):e.__proto__=t)}(t,i.Component),r(t,[{key:"render",value:function(){return a.default.createElement(o.default,d)}}]),t}();t.default=p},function(e,t,n){"use strict";Object.defineProperty(t,"__esModule",{value:!0});var r=function(){function e(e,t){for(var n=0;n<t.length;n++){var r=t[n];r.enumerable=r.enumerable||!1,r.configurable=!0,"value"in r&&(r.writable=!0),Object.defineProperty(e,r.key,r)}}return function(t,n,r){return n&&e(t.prototype,n),r&&e(t,r),t}}(),i=n(0),a=s(i),o=s(n(4)),l=s(n(8));function s(e){return e&&e.__esModule?e:{default:e}}var u=a.default.createElement("div",null,a.default.createElement("div",null,"In order to achieve real time data analysis , when ACE detects the color-changing sample, it will send a message to the user. Between ACE and user’s mobile device, it needs a transfer protocol to communicate with each other. Here, we choose MQTT (Message Queuing Telemetry Transport) to upload the vibrio concentration onto the cloud. It is a lightweight messaging protocol for small sensors and mobile devices, optimized for high-latency or unreliable networks.  Therefore, this kind of method is useful in remote area where network bandwidth is limited, especially for the fish farm in Taiwan. We’ve designed an app for farmers to obtain information in time. In our app, we separate into three parts: Pond’s condition, Suggestions, and Multiple Pond map. "),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.software.app}))),c=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},"We’ve divided different bacteria’s concentration into three levels, Safe, Warning, and Danger. The standard of this division is refer to the advice of the Fisheries Research Center of Republic of China. After visiting the Marine Life museum Research center, the researcher also tell us that usually the concentration for vibrio to be pathogenic is >104CFU/ml."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.software.line})),a.default.createElement("div",{className:"part"},"Furthermore, in order to let the farmers keep track of how the vibrio in their farm has been growing, we’ve made a history record chart, of which the Y coordinate is concentration, and the X coordinate is time. The concentration points will be drawn every hour.  The chart can be drag, zoom in and out for a closer look. By knowing the condition of their pond beforehand and even predict how the amount of the vibrios might grow, the farmers can do precaution in order to prevent vibrios from harming their fish or shrimp.")),f=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},"When the concentration of vibrios is under the safety standard, the fisheries research center recommend the farmers to change the water environment, or adjust the amount of feed. Only when the condition is in danger that they can add the medicine.   "),a.default.createElement("div",{className:"part"},"In addition, we will send notification bar through android phones to farmers if the amount of vibrios exceed standard."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.software.warn}))),d=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},"The reason why we want to combine google map into our APP is that one farmer usually own more than one pond.  Then if they want to keep track of different pond, they need to number their farm. Most of the time, it’s difficult for farmers to memorize which pond does this number really referring to, which indicate that the managing system is not so user friendly. "),a.default.createElement("div",{className:"part"},"In our map, farmers can locate its position and to help its pond create a name.  Next time when they want to inspect the condition of each pond, they can just press on the name label on the map. In order to make the system more intuitive, we add in the function of turning the normal map into 3D map and satellite(RealView) one, which is more close to the real view on the ground."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.software.map}))),p={viewbar:{Title:"Software",Subtitle:"An app for real-time analysis of pond’s condition",image:o.default.image.topfull.software},content:{Introduction:u,parts:[{Title:"Pond’s condition",Description:c},{Title:"Suggestions",Description:f},{Title:"Multiple pond management map",Description:d}]}},h=function(e){function t(){return function(e,t){if(!(e instanceof t))throw new TypeError("Cannot call a class as a function")}(this,t),function(e,t){if(!e)throw new ReferenceError("this hasn't been initialised - super() hasn't been called");return!t||"object"!=typeof t&&"function"!=typeof t?e:t}(this,(t.__proto__||Object.getPrototypeOf(t)).apply(this,arguments))}return function(e,t){if("function"!=typeof t&&null!==t)throw new TypeError("Super expression must either be null or a function, not "+typeof t);e.prototype=Object.create(t&&t.prototype,{constructor:{value:e,enumerable:!1,writable:!0,configurable:!0}}),t&&(Object.setPrototypeOf?Object.setPrototypeOf(e,t):e.__proto__=t)}(t,i.Component),r(t,[{key:"render",value:function(){return a.default.createElement(l.default,p)}}]),t}();t.default=h},function(e,t,n){"use strict";Object.defineProperty(t,"__esModule",{value:!0});var r=function(){function e(e,t){for(var n=0;n<t.length;n++){var r=t[n];r.enumerable=r.enumerable||!1,r.configurable=!0,"value"in r&&(r.writable=!0),Object.defineProperty(e,r.key,r)}}return function(t,n,r){return n&&e(t.prototype,n),r&&e(t,r),t}}(),i=n(0),a=s(i),o=s(n(8)),l=s(n(4));function s(e){return e&&e.__esModule?e:{default:e}}var u=a.default.createElement("div",null,a.default.createElement("div",null,"The importance of damage caused by Vibrio would not be elucidated here. You can see the detailed information in",a.default.createElement("a",{href:"https://2018.igem.org/Description"}," Description"),"."),a.default.createElement("div",{className:"part"},a.default.createElement("div",null,"Nowadays, there are three possible scenarios for aquatic agriculture to face pathogenic Vibrio:"),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",null,"1. The infection of Vibrio is usually happening all of a sudden. So when fisherman aware of the infection, it often causes significant damage."),a.default.createElement("div",null,"2. The conventional Vibrio testing method is time-consuming and manpower-consuming."),a.default.createElement("div",null,"3. The current water quality testing equipment is mainly focusing on the physical and chemical index, there are no devices for testing bacteria"))),a.default.createElement("div",{className:"part"},a.default.createElement("div",null,"However, our automatic Vibrio-detecting system is able to solve these problems."),a.default.createElement("div",{style:{display:"flex",alignItems:"flex-end"}},a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:l.default.image.application.tcbs}),a.default.createElement("div",{className:"undertext"},"TCBS medium")),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:l.default.image.application.plate}),a.default.createElement("div",{className:"undertext"},"MV-kit"))))),c=a.default.createElement("div",null,a.default.createElement("div",null,"MV-kit was developed by Fisheries Research Institute in Taiwan. It’s a kind of selective medium for Vibrio and it can detect 17 different aquatic pathogenic Vibrio. Through observing the time of color-changing, we can know the approximate Vibrio concentration in the sample. After our modification, within 24 hours, the minimal detectable Vibrio concentration is 100 CFU/ mL. Take Vibrio harveyi for example:"),a.default.createElement("div",{style:{display:"flex",alignItems:"flex-end"}},a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:l.default.image.application.concentration}),a.default.createElement("div",{className:"undertext"},"CFU vs. O.D.")),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:l.default.image.application.time}),a.default.createElement("div",{className:"undertext"},"CFU vs. Time of color-changing"))),a.default.createElement("div",null,"We build a relationship of O.D. value and the amount of bacteria via spreading method. Then, using O.D. value as a reference point, we build another relationship between the color-changing time of MV-kit and the amount of bacteria. Accordingly, we can get a significant correlation of bacteria concentration and the color-changing time of MV-kit")),f=a.default.createElement("div",null,a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:l.default.image.application.flow})),a.default.createElement("div",{className:"part"},"We provide a complete service for the prevention of Vibrio-infection. Our ACE collects the water sample from the fish pond and drips it into the plate automatically. Simultaneously, the device provides a closed environment with constant temperature for the reaction. Also, the device monitors the color-changing time at all times. As long as the color changes and achieves the condition we have set up, the data will be recorded and be uploaded to the server. After analyzing, the final result will be transferred to the clients’ cellphone applications. If the concentration of Vibrio overtakes the minimal hazardous value which is 1000 CFU/ mL, the application will warn the fisherman."),a.default.createElement("div",{className:"part"},"Through our system, our clients can take control of the situation of their fish ponds and use our engineered E. coli or other plans to solve the problems.")),d=a.default.createElement("div",null,a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:l.default.image.application.stp})),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"1. Segmentation"),a.default.createElement("div",null,a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",{className:"part-title"},"Merchandises in current market have to face some difficulties:"),a.default.createElement("div",null,"The water quality detecting systems in the current market are merely focusing on the particular physical and chemical indexes in the waters, for example, pH value, redox potential, dissolved oxygen, etc. They determine the water quality by those indexes, however, the change of situation in the water is quite fast and complex. We cannot receive the correct and real-time information of Vibrio in the water. When we aware of the abnormality, the pathogenic Vibrio usually causes the death of fish and shrimps significantly.")),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",{className:"part-title"},"Long-term detection with high specificity:"),a.default.createElement("div",null,"The MV-kit developed by Fisheries Research Institute is able to detect the Vibrio in the water sample quantitatively and with high specificity. Our system can automatically monitor the change of Vibrio concentration over a long period of time. Our clients are able to notice the condition in the early stage of Vibrio growth and adopt the solutions.")),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",{className:"part-title"},"A real-time and automatic system:"),a.default.createElement("div",null,"Our mechanism design allows the device to collect the sample automatically and continuously. Furthermore, we also program an application which analyzes the Vibrio concentration automatically. Last but not least, we achieve the effect of real-time monitoring and report through IoT technology.")),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",{className:"part-title"},"Fast, simple, and low-cost:"),a.default.createElement("div",null,"The mechanism of our reaction is the simple color-changing process. Compared to the conventional Vibrio culture methods, our clients haven’t to equipped with the professional skills of bacteria culture, also, the process of testing becomes safer and faster. In comparison with the water quality testing system in the market, our system is low-cost and is able to express the Vibrio concentration directly and accurately."))),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:l.default.image.application.vs}))),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"2. Targeting"),a.default.createElement("div",null,a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",null,"Aquatic agriculture industry:"),a.default.createElement("div",null,"Aquatic agriculture is an important industry and it stimulates the economic growth in Taiwan every year. However, the Vibrio-related infection leads to inestimable economic damage locally and globally. Our system aims to provide real-time, long-term, and low-cost monitoring toward Vibrio. Through the automatic system, our clients can reduce the manpower-cost and ensure their fish ponds are safe at any time. In addition, our mechanism design is quite simple so that the maintenance cost is also quite low. The only thing fisherman has to do is change the consumable materials regularly.")),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",{className:"part-title"},"Suppliers of the water quality testing system:"),a.default.createElement("div",null,"Currently, there are several water quality testing system vendors in Taiwan, but there is no any company provides the automatic Vibrio-detecting system. Our device is easy to be integrated with other related systems to reduce the vendors’ research and development cost. In this way, we believe that we can establish a better platform and to create a win-win situation.")))),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"3. Positioning"),a.default.createElement("div",null,a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",{className:"part-title"},"Intelligent fishery:"),a.default.createElement("div",null,a.default.createElement("div",null,"With the advance of science and technology, the IoT popularizes gradually in current society. Besides, the traditional agriculture industries transform into the so-called “agriculture 4.0”. "),a.default.createElement("div",null,"Let’s take an in-depth look at aquatic agriculture in Taiwan. Our country is famous for its aquatic agriculture industry, however, the conventional way is quite time-consuming. Additionally, following with the global warming, the aquatic agriculture faces a huge challenge. In the train of those facts, the industrial population structure becomes imbalance and thus, exerting a crucial impact on the future of industrial development ultimately."),a.default.createElement("div",null,"In order to turn the tables, the concept and development of intelligent fishery are emerging in recent years. It leads our traditional fishery industry into a new era, raising the operational efficacy, the income, and reducing the cost. Based on the concept, our system is expected to solve the Vibrio-monitoring problems and to establish a database to find an optimized solution."))),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",{className:"part-title"},"Future resolution:"),a.default.createElement("div",null,"Hopefully, we can create a complete detecting platform for different pathogenic bacteria in the future. With low-cost and high-efficiency, our platform is able to minimize the economic damage result from the bacterial infection."))))),p={viewbar:{Title:"Application",Subtitle:"Automatic Vibrio-detecting System",image:l.default.image.topfull.application},content:{Introduction:u,parts:[{Title:"MV-kit",Description:c},{Title:"Automatic Vibrio-detecting System",Description:f},{Title:"Commercial Analysis",Description:d}]}},h=function(e){function t(){return function(e,t){if(!(e instanceof t))throw new TypeError("Cannot call a class as a function")}(this,t),function(e,t){if(!e)throw new ReferenceError("this hasn't been initialised - super() hasn't been called");return!t||"object"!=typeof t&&"function"!=typeof t?e:t}(this,(t.__proto__||Object.getPrototypeOf(t)).apply(this,arguments))}return function(e,t){if("function"!=typeof t&&null!==t)throw new TypeError("Super expression must either be null or a function, not "+typeof t);e.prototype=Object.create(t&&t.prototype,{constructor:{value:e,enumerable:!1,writable:!0,configurable:!0}}),t&&(Object.setPrototypeOf?Object.setPrototypeOf(e,t):e.__proto__=t)}(t,i.Component),r(t,[{key:"render",value:function(){return a.default.createElement(o.default,p)}}]),t}();t.default=h},function(e,t,n){"use strict";Object.defineProperty(t,"__esModule",{value:!0});var r=function(){function e(e,t){for(var n=0;n<t.length;n++){var r=t[n];r.enumerable=r.enumerable||!1,r.configurable=!0,"value"in r&&(r.writable=!0),Object.defineProperty(e,r.key,r)}}return function(t,n,r){return n&&e(t.prototype,n),r&&e(t,r),t}}(),i=n(0),a=s(i),o=s(n(8)),l=s(n(4));function s(e){return e&&e.__esModule?e:{default:e}}var u=a.default.createElement("div",null,a.default.createElement("div",null,"In order to control Vibrio cell-population density and maintain a bio-friendly environment in a sustainable way, we introduce the concept of biocontrol, which is known as the prey-predator model, to our system. We create a system that can automatically detect, compare, and control the cell-population density. The system is composed of three parts, which are a detector, comparator, and controller respectively (Fig 1). These three parts are connected together in the same gene circuit and function sequentially so that the concentration of Vibrio in the surroundings can be kept in a specific range. Then, a succession model can be achieved."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:l.default.image.wetlab_overview.system}),a.default.createElement("div",{className:"undertext"},"Fig 1. Component of cell population controlling system."))),c=a.default.createElement("div",null,"Our gene circuit design can be divided into three parts described down below."),f=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},"The Detector module is responsible for detecting the concentration of Vibrio species in the surroundings. We found that most of pathogenic Vibrio use quorum sensing to communicate with other individuals. Lux is one of the Vibrio quorum sensing systems. With Lux system, the Vibrio has the ability to secrete and detect specific chemicals, acyl homoserine lactones (AHLs, Fig2). Signal transduction relies on AHLs, and bacteria respond to it with different gene expression."),a.default.createElement("div",{className:"part"},"We use Lux gene originally isolated from Vibrio fischeri as our AHL sensor. The gene system is composed of LuxI, LuxR, and pLux. LuxI is a synthase that converts S-Adenosyl methionine (SAM) into one of specific AHLs, 3OC6HSL (Fig2). LuxR is an activating protein, and it will stimulate the transcription from Lux promoter (pLux) when 3OC6HSL binds to it.[1] We modify LuxI with RFP reporter to the E. coli. playing the role of Vibrio. We also modify LuxR and pLux into our genetically engineered E. coli., the cell-population controlling system, so that AHL level in the environment can be detected and the Vibrio population can be known (Fig3).        "),a.default.createElement("div",{className:"illustration"},a.default.createElement("div",{className:"illust-image-wrapper"},a.default.createElement("img",{className:"illust-image",src:l.default.image.wetlab_overview.ahl})),a.default.createElement("div",{className:"undertext"},"Fig 2. Chemical structure of 3-oxo-C6-HSL.")),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:l.default.image.wetlab_overview.detection}),a.default.createElement("div",{className:"undertext"},"Fig 3. An AHL detection scheme showing that AHL-LuxR complex activates the gene downstream from pLux."))),d=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},"The comparator module is responsible for comparing the cell population density of Vibrio with our engineered E. coli. to see if it exceeds the pathogenic level. It receives information getting from the detector module. If the concentration of Vibrio is higher than engineered E. coli., which means LuxR-AHL complex is present, the anti-Vibrio gene will be transcribed. On the other hand, if the concentration of engineered E. coli. is higher than Vibrio, which means LuxR-AHL complex is absent, the suicide gene will be transcribed to kill E. coli. itself. "),a.default.createElement("div",{className:"part"},"Our initial effort is to design a gene circuit contain LacR to accomplish our goal. However, it turns out that there are some problems and drawbacks. While AHL presences, LacR should be produced and bound to pLac promoter to inhibit suicide gene being expressed. Yet, LacR can’t 100 % bind to pLac and this happens in most operons, so-called leakage problem. Suicide gene may be expressed, and E. coli. will suicide even at high AHL concentration. This would cause the system to be collapsed, and the result became out of our expectations (Fig 4)."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:l.default.image.wetlab_overview.lacl}),a.default.createElement("div",{className:"undertext"},"Fig 4. Our initial design using Lac operon to regulate genes expression.")),a.default.createElement("div",{className:"part"},"We come up with a better solution for our kill switch by using riboswitch and Small Transcriptional-Activating RNA (STAR) to regulate our gene. STAR is a small RNA with its sequence completely complementary to riboswitch. Riboswitch is an intrinsic transcription terminator, a DNA segment with special secondary structure. It forms a hairpin structure and inhibits transcriptions of the genes downstream.[2] When STAR binds to the riboswitch, it will be turned on (Fig 5, 6). Current studies show that riboswitch can regulate protein better than operon because of the higher affinity between STAR and riboswitch. In addition, STAR is smaller than regular proteins and it degrades faster than regular protein, therefore can respond to environmental change more rapidly than operon."),a.default.createElement("div",{className:"illustration"},a.default.createElement("div",{className:"illust-image-wrapper"},a.default.createElement("img",{className:"illust-image",src:l.default.image.wetlab_overview.riboswitch})),a.default.createElement("div",{className:"undertext"},"Fig 5. The secondary structure of riboswitch and STAR")),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:l.default.image.wetlab_overview.star}),a.default.createElement("div",{className:"undertext"},"Fig 6. STAR turns on the target and activates the downstream transcriptional process")),a.default.createElement("div",{className:"part"},"We also use a small RNA segment called anti-STAR in our design. Its sequence is totally complementary to STAR, but has higher affinity than riboswitch does. When anti-SATR exists, STAR will bind to it prior to riboswitch. Consequently, when Lux-AHL triggers the transcription of anti-STAR, it binds to STAR and the riboswitch is turned off (Fig 7). When Lux-AHL is absent and anti-STAR isn’t generated, STAR will turn on the riboswitch."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:l.default.image.wetlab_overview.anti}),a.default.createElement("div",{className:"undertext"},"Fig 7. Anti-STAR makes STAR lose it functionality. The gene downstream is still inhibited by the target."))),p=a.default.createElement("div",null,a.default.createElement("div",null,"We use two growth inhibitors to control populations of Vibrio and engineered E. coli. One is for inhibiting Vibrio, and another is for inhibiting engineered E. coli. At first, we would like to use anti-Vibrio substances in our design. However, our lab belongs to Bio-safety Level 1 (BSL1) and we can’t use real Vibrio in our experiments to test the toxicity of anti-Vibrio substance due to bio-safety concern. Therefore, we use colicin M to kill E. coli. (fake Vibrio) in order to prove our bio-control model. On the other hand, we use toxin small RNA (tsRNA) and endolysin as suicide substances to kill engineered E. coli. itself."),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"1. Colicin M"),a.default.createElement("div",null,"Colicin M blocks the biosynthesis of peptidoglycan (murein) and O-antigen. Colicin M contains activity (cma) and immune gene (cmi). The product of cmi is an immune protein, preventing colicin M to inhibit cell wall synthesis by binding to and thus neutralizing it. In addition, most colicins are released by autolysis, but colicin M lacks a lysis gene.[3-4] It has less tendency to cause the death of producer cells, so it’s a suitable protein for the purpose of killing other bacteria without harming producer cells.")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"2. Toxic small RNA (tsRNA)"),a.default.createElement("div",null,"Toxic small RNAs are small non-coding RNA. We use the tsRNA isolated from Burkholderia cenocepacia. The Current study showed that it is toxic to the E. coli. host and reproducibly inhibit E. coli growth. TsRNA harbored a sequence that was complementary to the consensus ribosomal binding site (RBS). It is hypothesized that it might target and misregulate essential genes within the E. coli cell. TsRNA cannot penetrate the bacterial cell from the outside, so it is unable to affect bacteria except E. coli. host cells.[5] Additionally, producing tsRNA doesn’t require a translational process, thus the response time is more rapid. As a result, tsRNA become an ideal suicide substance for our design.")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"3. Endolysin"),a.default.createElement("div",null,"Endolysin is a peptidoglycan hydrolase that is secreted by double-stranded DNA lambda phage to comprise the bacterial cell wall. It causes cell lysis by disrupting murein structure.")),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:l.default.image.wetlab_overview.mechanism}),a.default.createElement("div",{className:"undertext"},"Fig 8. The overall gene circuit design"))),h=a.default.createElement("div",null,a.default.createElement("div",{className:"innerpart"},"1. Visick, K. L.; Foster, J.; Doino, J.; McFall-Ngai, M.; Ruby, E. G., Vibrio fischeri lux Genes Play an Important Role in Colonization and Development of the Host Light Organ. Journal of Bacteriology 2000, 182 (16), 4578-4586."),a.default.createElement("div",{className:"innerpart"},"2. Chappell, J.; Takahashi, M. K.; Lucks, J. B., Creating small transcription activating RNAs. Nature chemical biology 2015, 11 (3), 214-20."),a.default.createElement("div",{className:"innerpart"},"3. Harkness, R. E.; Olschlager, T., The biology of colicin M. FEMS microbiology reviews 1991, 8 (1), 27-41."),a.default.createElement("div",{className:"innerpart"},"4. Zeth, K.; Römer, C.; Patzer, S. I.; Braun, V., Crystal Structure of Colicin M, a Novel Phosphatase Specifically Imported by Escherichia coli>. The Journal of Biological Chemistry 2008, 283 (37), 25324-25331."),a.default.createElement("div",{className:"innerpart"},"5. Kimelman, A.; Levy, A.; Sberro, H.; Kidron, S.; Leavitt, A.; Amitai, G.; Yoder-Himes, D. R.; Wurtzel, O.; Zhu, Y.; Rubin, E. M.; Sorek, R., A vast collection of microbial genes that are toxic to bacteria. Genome research 2012, 22 (4), 802-9.")),m={viewbar:{Title:"Wetlab",Subtitle:"Detector, comparator, and controller module",image:l.default.image.topfull.wetlab_overview},content:{Introduction:u,parts:[{Title:"Cell Population Controlling System",Description:c},{Title:"Detector",Description:f},{Title:"Comparator",Description:d},{Title:"Controller",Description:p},{Title:"Reference",Description:h}]}},g=function(e){function t(){return function(e,t){if(!(e instanceof t))throw new TypeError("Cannot call a class as a function")}(this,t),function(e,t){if(!e)throw new ReferenceError("this hasn't been initialised - super() hasn't been called");return!t||"object"!=typeof t&&"function"!=typeof t?e:t}(this,(t.__proto__||Object.getPrototypeOf(t)).apply(this,arguments))}return function(e,t){if("function"!=typeof t&&null!==t)throw new TypeError("Super expression must either be null or a function, not "+typeof t);e.prototype=Object.create(t&&t.prototype,{constructor:{value:e,enumerable:!1,writable:!0,configurable:!0}}),t&&(Object.setPrototypeOf?Object.setPrototypeOf(e,t):e.__proto__=t)}(t,i.Component),r(t,[{key:"render",value:function(){return a.default.createElement(o.default,m)}}]),t}();t.default=g},function(e,t,n){"use strict";Object.defineProperty(t,"__esModule",{value:!0});var r=function(){function e(e,t){for(var n=0;n<t.length;n++){var r=t[n];r.enumerable=r.enumerable||!1,r.configurable=!0,"value"in r&&(r.writable=!0),Object.defineProperty(e,r.key,r)}}return function(t,n,r){return n&&e(t.prototype,n),r&&e(t,r),t}}(),i=n(0),a=u(i),o=u(n(4)),l=u(n(8)),s=u(n(136));function u(e){return e&&e.__esModule?e:{default:e}}var c=a.default.createElement("div",null,a.default.createElement("div",{className:"illustration"},a.default.createElement("div",{style:{margin:"auto",width:"500px"}},a.default.createElement("img",{className:"illust-image",src:o.default.image.model.interaction}))),a.default.createElement("div",{className:"part"},"We genetically modify the E.coli, giving them the ability to detect Vibrio concentration, and secrete some compound to supress Vibrio and itself. "),a.default.createElement("div",{className:"part"},"If the AHL concentration exceed a specific threshold, engineered E.coli will secrete antibacterial peptide Colisin M to kill Vibrio. Otherwise, the Vibrio concentration below the E.coli's detection threshold, engineered Ecoli will secrete tsRNA to commit suicide.")),f=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},a.default.createElement("div",null,"We revise this model from Lotka-Volterra model with logistic population groth, and the decay rate of compound."),a.default.createElement("div",{style:{width:"fit-content",margin:"auto"}},a.default.createElement(s.default.Provider,null,a.default.createElement("div",{style:{marginTop:"20px"}},a.default.createElement(s.default.Node,{inline:!0,formula:"{{dV} \\over{dt}} = r_v V - {{r_v} \\over {V_{max}}} V^2 -a VN"})),a.default.createElement("div",{style:{marginTop:"20px"}},a.default.createElement(s.default.Node,{inline:!0,formula:"{{dA} \\over {dt}} = k_AV - \\lambda_A A"})),a.default.createElement("div",{style:{marginTop:"20px"}},a.default.createElement(s.default.Node,{inline:!0,formula:"{{dE} \\over {dt}} = bEA - cES - \\lambda_E E"})),a.default.createElement("div",{style:{marginTop:"20px"}},a.default.createElement(s.default.Node,{inline:!0,formula:"{{dC} \\over {dt}} = k_CE - \\lambda_C C "})),a.default.createElement("div",{style:{marginTop:"20px"}},a.default.createElement(s.default.Node,{inline:!0,formula:"{{dS} \\over {dt}} = k_SE - \\lambda_S S"}))))),a.default.createElement("div",{className:"part"},"With the contraint of AHL threshold",a.default.createElement("div",{style:{width:"fit-content",margin:"auto"}},a.default.createElement(s.default.Provider,null,a.default.createElement("div",{style:{marginTop:"20px"}},a.default.createElement(s.default.Node,{inline:!0,formula:"A \\lt A_t, \\ k_C=0"})),a.default.createElement("div",{style:{marginTop:"20px"}},a.default.createElement(s.default.Node,{inline:!0,formula:"A \\geq A_t, \\ k_S=0"})))))),d=a.default.createElement("div",null,"With changing specific parameter, such as binding strength of antibacterial Colisin with Vibrio, binding strength of Ahl with Ecoli, and the detection threshold of AHL for engineered E.coli. We are able to simulate the population of two bacteria in oscillating, and a parameter sensitive constant mode.",a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.model.result}),a.default.createElement("div",{className:"undertext"},"Two figure are just a slight change of the Binding strength of Ahl with Ecoli"))),p=a.default.createElement("div",null,"Code detail On ",a.default.createElement("a",{href:"https://github.com/igem-tw-nthu/model-2018"},"Github")),h={viewbar:{Title:"Model",Subtitle:"Build a simple bio-oscillator",image:o.default.image.topfull.model},content:{Introduction:"This year 2018 iGEM NTHU_Taiwan aims to solve the Vibrio infection in aquaculture with bio-competiotion. We are inspired by the peroidic pattern of Lotka-Volterra model, and try to control the Vibrio population with engineered E.coli.",parts:[{Title:"Interaction",Description:c},{Title:"Equations",Description:f},{Title:"Discussion",Description:d},{Title:"Soucre Code",Description:p}]}},m=function(e){function t(){return function(e,t){if(!(e instanceof t))throw new TypeError("Cannot call a class as a function")}(this,t),function(e,t){if(!e)throw new ReferenceError("this hasn't been initialised - super() hasn't been called");return!t||"object"!=typeof t&&"function"!=typeof t?e:t}(this,(t.__proto__||Object.getPrototypeOf(t)).apply(this,arguments))}return function(e,t){if("function"!=typeof t&&null!==t)throw new TypeError("Super expression must either be null or a function, not "+typeof t);e.prototype=Object.create(t&&t.prototype,{constructor:{value:e,enumerable:!1,writable:!0,configurable:!0}}),t&&(Object.setPrototypeOf?Object.setPrototypeOf(e,t):e.__proto__=t)}(t,i.Component),r(t,[{key:"render",value:function(){return a.default.createElement(l.default,h)}}]),t}();t.default=m},function(e,t,n){"use strict";Object.defineProperty(t,"__esModule",{value:!0}),t.default=void 0;var r=function(){function e(e,t){for(var n=0;n<t.length;n++){var r=t[n];r.enumerable=r.enumerable||!1,r.configurable=!0,"value"in r&&(r.writable=!0),Object.defineProperty(e,r.key,r)}}return function(t,n,r){return n&&e(t.prototype,n),r&&e(t,r),t}}(),i=n(0),a=s(i),o=s(n(4)),l=s(n(8));function s(e){return e&&e.__esModule?e:{default:e}}var u=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Transformation"),a.default.createElement("div",null,"During this summer, we had done lots of digestion-ligation and transformation experiments! We enjoyed and had fun exploring the mystery of synthetic biology!"),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.experiment.transform}),a.default.createElement("div",{className:"undertext"},"Fig. 1: Successful transformation of part BBa_K2774008 (Left) and BBa_K2774005 (Right)")),a.default.createElement("div",null,"Utilizing iGEM distribution kit and parts from the registry, we assembled various bio-bricks to construct our genes. We constructed our part, BBa_K2774008, by using pLac with RBS (BBa_J04500), endolysin (BBa_K1378032) with GFP (E0240). The left LB plate was the transformation of BBa_K2774008. The transformation was success and therefore some of the colony emits GFP fluorescence! And the right LB plate was the transformation of part BBa_K2774005, the RFP shows the right colony with successful ligation plasmid. In addition, thanks for the established technology of IDT (Integrated DNA Technologies), providing us the gene fragments. Finally, we successfully constructed tsRNA (BBa_K2774001).")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Electrophoretogram"),a.default.createElement("div",null,"Here are the parts we assemble successfully. We use VF2 and VR primer to check our gene’s length, all of them are on pSB1C3."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.experiment.elec}),a.default.createElement("div",{className:"undertext"},"Fig.2: Electrophoretogram of our parts which we had submitted to registry, all of them show properly length (Using VF2 and VR primer, the length will increase about 300 bp)")),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.experiment.table_1}),a.default.createElement("div",{className:"undertext"},"Table 1: This table’s genes correspond to each lane of Fig. 2 from left to right.")))),c=a.default.createElement("div",null,"We use TCBS (Thiosulfate-citrate-bile salts-sucrose agar, a type of selective agar culture plate that is used in microbiology laboratories to isolate Vibrio) to examine Vibrio’s concentration in fish farm. Our HP member and wet lab member visit the fish farm in Taiwan, Chiayi. We collected the water sample from different pool, and sent it back to the lab to culture the sample on TCBS.",a.default.createElement("div",{className:"illustration"},a.default.createElement("div",{className:"illust-image-wrapper",style:{width:"600px"}},a.default.createElement("img",{className:"illust-image",src:o.default.image.experiment.sample})),a.default.createElement("div",{className:"undertext"},"Fig.3 : Collection sample from traditional fish farm in Taiwan")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Result"),a.default.createElement("div",null,"Sample from fish farm shown positive result, we found there were colony appeared on TCBS plate. Implying that Vibrio indeed exist in fish farm, and may infect shrimp or other marine creatures. In addition, the colony color stands for different types of Vibrio:"),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",null,"Vibrio Cholerae: Large yellow colonies. "),a.default.createElement("div",null,"Vibrio harveyi / Vibrio fischeri: Greyish-green to bluish-green colonies which show luminescence in dark.")),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:o.default.image.experiment.vibrio}),a.default.createElement("div",{className:"undertext"},"Fig.4 : Testing Sample from fish farm on TCBS"),a.default.createElement("div",{className:"undertext"},"Right plate shows Vibrio harveyi and the left pate shows Vibrio Cholerae")))),f=a.default.createElement("div",null,"Our detection system is designed to make E. coli to obtain the ability to sense the concentration of Vibrio in the environment. We found that most of the pathogenic Vibrio use “quorum sensing” to communicate with each individual and to express different genes. Quorum sensing, a system which is able to control the gene expression followed by the population density. The signal transmission of quorum sensing is totally depending on the auto-inducer, acyl homoserine lactones (AHL), a small chemical molecule which is produced by bacterial enzyme (Fig. 5). Moreover, the concentration of AHL is directly proportional to bacteria density. When AHL concentration exceeds the threshold, it will bind with the protein (LuxR) and trigger the promoter (pLUX) to express the downstream genes.",a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:o.default.image.experiment.ahl}),a.default.createElement("div",{className:"undertext"},"Fig. 5: The schematic of our gene circuits")),a.default.createElement("div",{className:"illustration",style:{width:"300px"}},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:o.default.image.experiment.structure}),a.default.createElement("div",{className:"undertext"},"Fig. 6: The structure of AHL")),"Our detection system is using Lux system. When the Vibrio density is high, the AHL concentration in the environment is high. Hence, our engineered E. coli can detect the Vibrio density and can release the anti-Vibrio substances. (Fig. 7)",a.default.createElement("div",{className:"illustration"},a.default.createElement("div",{className:"illust-image-wrapper",style:{width:"500px"}},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:o.default.image.experiment.cycle})),a.default.createElement("div",{className:"undertext"},"Fig. 7: The schematic of our detection system")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Exp. 1: AHL Sensor with Quantitative AHL "),a.default.createElement("div",null,"We constructed our AHL detector (BBa_K2774000), which can produce LuxR consistently by pLac promoter and can express GFP while LuxR combine with AHL and trigger pLUX (R0062). While AHL presents, AHL and LuxR will form a complex and trigger pLUX to express GFP. (Fig. 8)"),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.experiment.sensor}),a.default.createElement("div",{className:"undertext"},"Fig. 8: AHL Bio-Detector")),a.default.createElement("div",null,"In order to test our gene’s function, we added different concentration of AHL (3-oxo-C6-HSL, synthesized by Sigma) to DH5α cells transformed with BBa_K2774000, and measured GFP expression hourly."),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Procedure:"),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",{className:"innerpart"},"1. Prepare DH5α cells which transformed with BBa_K2774000 and liquid culture in LB overnight."),a.default.createElement("div",{className:"innerpart"},"2. Diluted eight bacterial culture samples in fresh LB (50 μL O/N culture with 4950 μL fresh LB)"),a.default.createElement("div",{className:"innerpart"},"3. Pre-culture until O.D.600 reached 0.4, making sure bacteria is growing in log phase (better expression capability) and add IPTG (1mM) to induce the expression of LuxR."),a.default.createElement("div",{className:"innerpart"},"4. Add different amount of AHL into different samples (0.01, 0.1, 0.5, 1, 5, 10, 100 nM) and 0 M AHL for positive control."),a.default.createElement("div",{className:"innerpart"},"5. Measure the O.D.600 value and expression of GFP (λexcitation = 490 nm; λemission = 530 nm) hourly.")),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.experiment.exp_1}),a.default.createElement("div",{className:"undertext"},"Fig. 9: GFP expression from different conc. of AHL"))),a.default.createElement("div",{className:"part"}),a.default.createElement("div",{className:"part-title"},"Result:"),a.default.createElement("div",null,"As the chart shows, we can find that AHL induced GFP expression as we expected. As the AHL concentration increased, the GFP expression increased until saturation. Besides, the optimized AHL concentration of inducing BBa_K2774000 is about 0.5-10 nM. Moreover, we found that if the concentration of AHL exceeds 1000 nM, it will inhibit BBa_K2774000 GFP expression."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:o.default.image.experiment.data_1}),a.default.createElement("div",{className:"undertext"},"Fig. 10: GFP expression from different conc. of AHL"))),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Exp. 2 : AHL sensor co-culture with fake Vibrio test"),"Since we cannot conduct the experiment related to Vibrio in our lab, we have to construct a fake Vibrio (by our engineered E. coli), which can also secrete AHL as Vibrio does. Therefore, we created our “pLac AHL generator” (BBa_2774004) by assembling “pLac + B0034” (BBa_J04500) and “LuxI” (BBa_C0161) together. Moreover, we added RFP to detect the population of fake vibrio (BBa_2774005)",a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Procedure:"),a.default.createElement("div",{className:"innerpart"}," 1. Prepare DH5α cells which transformed with BBa_2774000 (AHL sensor), BBa_2774004 (pLac AHL generator), and BBa_2774005 (pLac AHL generator + RFP) and liquid culture in LB overnight."),a.default.createElement("div",{className:"innerpart"}," 2. Diluted eight overnight bacterial culture in fresh LB with 1mM IPTG (50μL O/N culture with 4950 μL fresh LB)."),a.default.createElement("div",{className:"innerpart"}," 3. Pre-culture until O.D.600 reaches 0.4, making sure bacteria is growing in log phase (better expression capability)."),a.default.createElement("div",{className:"innerpart"}," 4. Mix different volume ratio of:"),a.default.createElement("div",null," BBa_2774004 (pLac AHL generator) with BBa_2774000 (AHL sensor)    "),a.default.createElement("div",null," BBa_2774005 (pLac AHL generator + RFP) with BBa_2774000 (AHL sensor)"),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:o.default.image.experiment.table_2})),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:o.default.image.experiment.table_3})),a.default.createElement("div",null,"5. Measure O.D.600, GFP and RFP expression after overnight culture.")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Result:"),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"a. BBa_2774005 (pLac AHL generator + RFP) co-culturing with BBa_2774000 (AHL sensor)"),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:o.default.image.experiment.exp_2}),a.default.createElement("div",{className:"undertext"},"Fig 11. AHL sensor with increasing concentration of AHL generator + RFP (from the left tube to the right tube). We centrifuged culture tubes in the left picture to get the ones in the right picture.")),a.default.createElement("div",{className:"innerpart"},"We co-cultured AHL sensor with increasing concentration of AHL generator + RFP. The picture indicates that AHL was indeed generated. When the two bacteria were mixed together, red and green fluorescence were both emitted and producing different colors.")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"b. BBa_2774004 (pLac AHL generator) co-culturing with BBa_2774000 (AHL sensor) "),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:o.default.image.experiment.table_4})),a.default.createElement("div",{className:"innerpart"},"In this experiment, we co-cultured AHL sensor with increasing concentration of AHL generator. We discovered that all liquid culture emitted green fluorescence except the one without adding AHL generator. We successfully proved that BBa_2774004 can express LuxI (AHL synthase) and generate AHL. We proposed that AHL concentration is linearly proportional to fake Vibrio concentration. However, the fluorescence intensities were almost the same among samples with increasing concentrations of AHL. We failed to construct a rational relation between fake Vibrio density and AHL concentration. We inferred the reason for this observation is that the rate of AHL synthesis is much higher than the rate of AHL degradation. The LuxI we used was the one without LVA-tag, so LuxI didn’t degrade instantly. In addition, we left the liquid culture overnight before measuring its fluorescence. Consequently, AHL was over-generated and the concentration of AHL was beyond the detection limit of AHL sensor.")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Exp. 3: Culture AHL sensor with Fake Vibrio’s supernatant")),a.default.createElement("div",null,"During Exp. 2 we find that overnight culturing and strong promoter cause excess AHL, which will not show the evidence of AHL concentration proportion to fake Vibrio concentration. Therefore we try a different way to experiment, we use weaker promoter and measure AHL concentration by AHL bio-sensor every hourly. Using interlab data we can get bacteria concentration by measuring OD600, and meanwhile we can centrifuge the tube to get supernatant which contain AHL secrete by the AHL generator that time."),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Procedure:"),a.default.createElement("div",{className:"innerpart"},"1. Prepare DH5α cells which transformed with BBa_2774000 (AHL sensor), BBa_2774002 (J23119+LxuI), and liquid culture in LB overnight."),a.default.createElement("div",{className:"innerpart"},"2.  Culture BBa_2774002 at microcentrifuge tube with (50μL O/N culture with 4950 μL fresh LB)."),a.default.createElement("div",{className:"innerpart"},"3. Measure microcentrifuge’s OD600 two hour and meanwhile centrifuge the tube to get supernatant contain AHL. Transfer the OD600 data to bacteria density by using inter lab data."),a.default.createElement("div",{className:"innerpart"},"4. Move 2.5 ml of supernatant contain AHL and 2.5ml fresh LB in new tube, and add AHL sensor simultaneously."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:o.default.image.experiment.exp_3}),a.default.createElement("div",{className:"undertext"},"Fig 12. The procedure of experiment 3."))),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Result:"),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:o.default.image.experiment.result_4}),a.default.createElement("div",{className:"undertext"},"Fig 13. AHL sensor with increasing concentration of AHL generator supernant")),a.default.createElement("div",{className:"innerpart"},"We transferred the supernatant from generator liquid culture which contained AHL into AHL sensor. The result showed that supernatant from a higher concentration of generator inducing AHL sensor to express higher amount of GFP. Moreover, after we transform the data and take the logarithm of generator density, we find a similar result which we had done in experiment 1. (Fig10)"),a.default.createElement("div",{className:"innerpart"},"Comparing exp1 and exp3 result, confirming that Vibrio density is proportional to the AHL concentration, at the condition of the real-time situation to avoid AHL accumulation. ")),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Conclusion:"),a.default.createElement("div",null,"We have successfully created a bio-sensor which can detect and react to different concentration of AHL. Moreover, our fake Vibrio (BBa_2774004) can also secrete AHL. We also confirmed that the Vibrio density is proportional to the AHL concentration. Our future work is to exchange gene which is under the control of pLux from GFP to anti-Vibrio substances."))))),d=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"innerpart"},"Based on our gene circuit, we need two kinds of substances to fulfill our goal. The first one is the suicide gene. When Vibrio concentration is low, our E. coli need to suicide themselves to maintain the equilibrium. Therefore, we introduced “tsRNA” (toxic small RNA), which was studied and experimented by Dr. Rotem . “tsRNA” was first found in Burkholderia cenocepacia and showed its toxic to E. coli. to inhibit E. coli growth. Moreover, references show that “tsRNA” couldn’t penetrate the bacterial cell wall, either from outside or inside. Therefore, it becomes the best candidate for our suicide gene. Although the mechanism of “tsRNA” is not clearly known, Dr. Rotem had tried complement sequences of “tsRNA” shown negative results. They hypothesized that “tsRNA” might target and mis-regulate essential genes within the E. coli cell. "),a.default.createElement("div",{className:"innerpart"},"To test the function of tsRNA for our suicide gene, we put tsRNA sequence under pLac promoter. Using IDT synthesized technology, we got the gene fragment (BBa_2774001)."),a.default.createElement("div",{style:{width:"300px"},className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:o.default.image.experiment.tsrna}),a.default.createElement("div",{className:"undertext"},"Fig 14. The secondary structure of tsRNA"))),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Procedure"),a.default.createElement("div",{className:"innerpart"},"1. Prepare DH5α which transformed with BBa_K2774001 and liquid culture in LB overnight."),a.default.createElement("div",{className:"innerpart"},"2. Diluted the overnight bacterial culture in fresh LB (50 μL O/N culture with 4950 μL fresh LB) "),a.default.createElement("div",{className:"innerpart"},"3. Pre-culture until O.D.600 reaches 0.2, making sure bacteria is growing in log phase (better expression capability) "),a.default.createElement("div",{className:"innerpart"},"4. Add IPTG (1mM) to induce tsRNA expression. (Adding ampicillin for positive control)"),a.default.createElement("div",{className:"innerpart"},"5. Measure O.D.600 hourly."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:o.default.image.experiment.tsrna_test}))),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Result"),a.default.createElement("div",{className:"innerpart"},"We induced tsRNA expression with IPTG at O.D.600 = 0.2. The result shows that tsRNA slightly inhibits E. coli growth, approximately 12%. We tested our BBa_K2774001 lots of times, but it still goes to the same result. Feeling confused, so we consulted Dr. Rotem for our situation, and got a generous reply:"),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:o.default.image.experiment.email})),a.default.createElement("div",{className:"innerpart"},"Dr. Rotem suggested us to check if our tsRNA mutated during transforming because of the inferiority of growth rate. Or to check tsRNA expression by using Northern blotting to check the RNA expression. Therefore, we sequenced our plasmid (tsRNA) and BLAST the result with our designed sequence: showing no mutation on tsRNA section."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",style:{backgroundColor:"transparent"},src:o.default.image.experiment.strength}),a.default.createElement("div",{className:"undertext"},"Fig 15. Sequencing result of tsRNA")),a.default.createElement("div",{className:"part-title"},"Conclusion"),a.default.createElement("div",{className:"innerpart"},"Our experiment result shows that the inhibition efficiency of tsRNA is 12%, which is significantly different from the one reported by Dr. Rotem. We sequenced our plasmid and no mutation was found. We inferred that the low inhibition efficiency is due to few expression of tsRNA. We believe this can be improved by using a stronger promoter to get a better result."))),p={viewbar:{Title:"Experiment",Subtitle:"",image:o.default.image.topfull.experiment},content:{Introduction:"In order to achieve our project’s goal, we constructed a few functional genes and tested their features. To sum up, our project can be divided into three parts. First of all is the “detection” part, we made our E. coli to detect the density of Vibrio and to respond. Then, it’s the part of “kill Switch”, we made our E. coli to switch from secreting anti-Vibrio substances to suicide to fulfilled bio-safety issues. Finally, it’s  “substances test” for anti-Vibrio and suicide substances which will be used in our project.",parts:[{Title:"Cloning",Description:u},{Title:"Testing Sample",Description:c},{Title:"Detection",Description:f},{Title:"Substances test",Description:d}]}},h=function(e){function t(e){return function(e,t){if(!(e instanceof t))throw new TypeError("Cannot call a class as a function")}(this,t),function(e,t){if(!e)throw new ReferenceError("this hasn't been initialised - super() hasn't been called");return!t||"object"!=typeof t&&"function"!=typeof t?e:t}(this,(t.__proto__||Object.getPrototypeOf(t)).call(this,e))}return function(e,t){if("function"!=typeof t&&null!==t)throw new TypeError("Super expression must either be null or a function, not "+typeof t);e.prototype=Object.create(t&&t.prototype,{constructor:{value:e,enumerable:!1,writable:!0,configurable:!0}}),t&&(Object.setPrototypeOf?Object.setPrototypeOf(e,t):e.__proto__=t)}(t,i.Component),r(t,[{key:"render",value:function(){return 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We both aimed to let students know more about iGEM and synthetic biology. During the camp, participants learned how to carry out simple experiments, e.g. plasmid extraction and transformation. We also shared with the students about our iGEM project and briefly introduced synthetic biology. The following are the schedule and all the material we used in the camp. About our files used in this camp are all available to",a.default.createElement("a",{href:"https://drive.google.com/drive/folders/19QhZWbbpJTddxGISAeXFJBAxcs_MuS_m?usp=sharing"}," download"),". The only thing you have to notice is the language, most are in Chinese."),f=a.default.createElement("div",null,a.default.createElement("div",{className:"illustration"},a.default.createElement("table",{className:"table"},a.default.createElement("tbody",null,a.default.createElement("tr",{className:"tablehead"},a.default.createElement("th",{className:"tablehead"}),a.default.createElement("th",{className:"tablehead"},"2018/08/10"),a.default.createElement("th",{className:"tablehead"},"2018/08/11")),a.default.createElement("tr",{className:"tablehead"},a.default.createElement("th",{className:"tablehead"},"9:00~12:00"),a.default.createElement("td",{className:"tabledata"},a.default.createElement("div",null,"iGEM Introduction & Team Project"),a.default.createElement("div",null,"Experiment Introduction "),a.default.createElement("div",null,"Pipetman Teaching ")),a.default.createElement("td",{className:"tabledata"},a.default.createElement("div",null,"Synthetic Biology Introduction"),a.default.createElement("div",null,"Experiment Introduction"),a.default.createElement("div",null,"Colony PCR"))),a.default.createElement("tr",{className:"tablehead"},a.default.createElement("th",{className:"tablehead"},"12:00~13:00"),a.default.createElement("td",{className:"tabledata"},"Lunch Time"),a.default.createElement("td",{className:"tabledata"},"Lunch Time")),a.default.createElement("tr",{className:"tablehead"},a.default.createElement("th",{className:"tablehead"},"13:00~16:00"),a.default.createElement("td",{className:"tabledata"},a.default.createElement("div",null,"Plasmid Extraction"),a.default.createElement("div",null,"Transformation"),a.default.createElement("div",null,"Bacteria Cultivation")),a.default.createElement("td",{className:"tabledata"},a.default.createElement("div",null,"Colony PCR"),a.default.createElement("div",null,"DNA Electrophoresis"),a.default.createElement("div",null,"Questionnaire Survey"))))))),d=a.default.createElement("div",null,a.default.createElement("div",null,"This year, 16 students joined our synthetic biology camp. The following are some feedback given by the students in the questionnaire survey. 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Or the students would be boring."),a.default.createElement("tr",{className:"tablehead"},a.default.createElement("td",{className:"tabledata"},"The contents of the experiments were quite funny, but the basic theories were difficult for high school students.")),a.default.createElement("tr",{className:"tablehead"},a.default.createElement("td",{className:"tabledata"},"Though I have not learned most of the terms about molecular biology experiments, the contents of experiments were interesting and inspiring which makes me acquire knowledge about molecular biology.")),a.default.createElement("tr",{className:"tablehead"},a.default.createElement("td",{className:"tabledata"},"If lecturers summarized all of the experiments to a table, it will be better!")),a.default.createElement("tr",{className:"tablehead"},a.default.createElement("td",{className:"tabledata"},"It’s my first time to do the molecular biology experiments and hopefully, you guys can achieve your goals!"))))))),p=a.default.createElement("div",null,a.default.createElement("div",null,"According to their feedback, we have achieved our goal: First, to introduce iGEM and synthetic biology. Second, teach high school students how to carry out synthetic biology experiment. Third, enhance their interest in genetic engineering. The synthetic biology camp this year is successful, and National Feng-Shan Senior High School are willing to cooperate with us next year! We hope our effort can encourage more students to do researches about genetic engineering, and maybe they will join iGEM soon!"),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:l.default.image.education.group}))),h={viewbar:{Title:"Education",Subtitle:u,image:l.default.image.topfull.education},content:{Introduction:"Joining iGEM allows us to carry out some interesting and creative experiments related to the latest researches. However, for ordinary senior high school students in Taiwan, they do not have the opportunity to engage in iGEM or the state-of-the-art knowledge of synthetic biology. Therefore, we decided to bring synthetic biology to high school students. We can not only tell them what synthetic biology is but also share our experience of joining iGEM as well as encourage them to participate next year!",parts:[{Title:"Summer Synthetic Biology Camp",Description:c},{Title:"Schedule",Description:f},{Title:"Feedback",Description:d},{Title:"Summary",Description:p}]}},m=function(e){function t(e){return function(e,t){if(!(e instanceof t))throw new TypeError("Cannot call a class as a function")}(this,t),function(e,t){if(!e)throw new ReferenceError("this hasn't been initialised - super() hasn't been called");return!t||"object"!=typeof t&&"function"!=typeof t?e:t}(this,(t.__proto__||Object.getPrototypeOf(t)).call(this,e))}return function(e,t){if("function"!=typeof t&&null!==t)throw new TypeError("Super expression must either be null or a function, not "+typeof t);e.prototype=Object.create(t&&t.prototype,{constructor:{value:e,enumerable:!1,writable:!0,configurable:!0}}),t&&(Object.setPrototypeOf?Object.setPrototypeOf(e,t):e.__proto__=t)}(t,i.Component),r(t,[{key:"render",value:function(){return a.default.createElement(o.default,h)}}]),t}();t.default=m},function(e,t,n){"use strict";Object.defineProperty(t,"__esModule",{value:!0});var r=function(){function e(e,t){for(var n=0;n<t.length;n++){var r=t[n];r.enumerable=r.enumerable||!1,r.configurable=!0,"value"in r&&(r.writable=!0),Object.defineProperty(e,r.key,r)}}return function(t,n,r){return n&&e(t.prototype,n),r&&e(t,r),t}}(),i=n(0),a=s(i),o=(s(n(10)),s(n(4))),l=s(n(8));function s(e){return e&&e.__esModule?e:{default:e}}var u=a.default.createElement("div",null,a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.interview.research})),a.default.createElement("div",null,"Fisheries Research Institute, COA, which is the government agency offers fish farmers to test the pathogen of fish and also, doing some researches on aquatic organisms as well. Deputy director of COA, C.I. Chang, who is a master in marine biology, recently did lots of researches on aquatic animal and pathogen, creating the MV-kit as well. The kit which function is to test the number of Vibrio in the water. And of course, FRI is the closest department between the local fish farmers and the public. As a result, we decided to visit him to get more helpful information and suggestions for our project. During our in-depth talk, we knew more about the Vibrio and, our core of the automated device,",a.default.createElement("a",{href:"https://2018.igem.org/Team:NTHU_Taiwan/Drylab-Overview"}," ACE"),". For example, after knowing the detailed information about the basic characteristics of Vibrio and the mechanism of MV-kit, making us want to combine the kit with our device and if it comes true, the device will be powerful. In the end, deputy director also gave us a few useful, potential advice and questions between our device and the kit. These feedbacks inspired us pretty much, such as whether our device would be influenced by the precipitation or not, or do we need some kinds of membrane to filter the aquatic sample or not, and in order to maintain the integrity and accuracy of our device, we have to test the efficiency of MV-kit in different environments and to find the simplest way that fish farmers can easily use. Nicely, the deputy director offering the plate of Vibrio harveyi and MV-kit for free in order to encourage us to make our experiment more complete.")),c=a.default.createElement("div",null,"In order to improve our project, we need to gain as much feedback as possible from the potential users; therefore, we’ve visited local fisherman in Taiwan, of which we can categorize into ecological and traditional breeding fish farmers."),f=a.default.createElement("div",null,a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.interview.tradition_1})),a.default.createElement("div",null,a.default.createElement("div",null,"In addition to visiting the ecological breeding farmers, we visited traditional fishermen in Kaohsiung and Chiayi, Mr. Chiu and Mr. Jian. They gave us some useful information, so we can improve our project more further."),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Mr. Chiu and Mr. Jian"),a.default.createElement("div",{className:"innerpart"},"Overall, they usually wait until the fish get sick and take them to the inspection center. If we can let them know about the condition of bacteria beforehand, they might take precautions, such as cleaning the water or adding the active powder to improve water quality and minimize the damage."),a.default.createElement("div",{className:"innerpart"},"When asking about what problem is the most crucial one they are facing, they’ve mentioned the uncertainty of electricity.  If the machine shut down or in a dangerous condition, sending a notification to farmers is essential. They also mentioned that adding the function of notifying farmer by message might be more practical than the smartphone application or the notification bar since farmer might be the elderly who do not use a smartphone. In light of their suggestions, we found the Chunghwa Telecom to seek for future collaboration. They told us that as long as we apply for a SIM card and a phone number, this kind of function can easily be done."),a.default.createElement("div",{className:"innerpart"},"To sum up, feedbacks and information are listed below. Plenty of them has greatly improved our projects:"),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",null,"1. Their working time is usually between 2 a.m. to 11 a.m., so they suggest us to test the sample during this time (since MV-kit needs 12~24 hr. to react completely)."),a.default.createElement("div",null,"2. They introduce the “TCBS” method: a common way to detect Vibrio sp. in traditional fish farm. This method need 1~2 day to complete."),a.default.createElement("div",null,"3. Add more features to ACE machine, e.g. water quality test (pH, NO3-, NO2-), other kit test (virus, fish parasite), and black out reminder."),a.default.createElement("div",null,"4. They recommend us to use text messages to be the notification of ACE machine. Because some of the traditional fishermen do not use smartphones."),a.default.createElement("div",null,"5. They recommend us to cooperate with fishermen who feed grouper or eel."),a.default.createElement("div",null,"6. As a traditional fisherman, they think our device has great potential."))))),d=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Mr. Tsai Hung-Jing’s Fish Farm"),a.default.createElement("div",null,a.default.createElement("div",{className:"innerpart"},"Before deciding which fish farm to visit, we looked for information on the Internet and finally found an interview published by CommonWealth Magazine, which is a famous magazine in Taiwan. The article showed that how successfully Mr. Tsai uses biological feed - water flea to take care of his shrimps and to make some profit. This idea of substituting chemical feed into water flea, an eco-friendlier one, matched the concept of iGEM. So, we called him and arranged the visit time without hesitation."),a.default.createElement("div",{className:"innerpart"},"Mr. Tsai was nice and friendly, he not only introduced his fish farm in such a detailed way but also answered all of our questions. He mentioned that the way he used to prevent Vibrio is to make the environment beneficial for good germs, which occupies the environment of the Vibrio, leaving no spaces for them to grow. He pointed out that people often uses ClO2 to kill Vibrio is unable to eradicate the problem completely. Once the pollutants are in, Vibrio will grow again. Consequently, we introduced our method to him and he sounded satisfied and looked forward to our final result. He also gave us some positive feedback toward our ACE. He said that if our service cost can be as low as 8,000 NT dollars, it will have a strong advantage and the market acceptance will be high since the price of the general detection devices need to be up to ten thousand NT dollars."),a.default.createElement("div",{className:"innerpart"},"The interview with Mr. Tsai really cheered us up, he let us understand the thing we do might help them in a practical way. Even though we went back to school, he still sent messages to encourage us.")),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.interview.eco_1}))),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Qiujia Aquaculture in Chiayi County"),a.default.createElement("div",null,a.default.createElement("div",{className:"innerpart"},"Qiujia Aquaculture has been established for 50 years, the main breed they sell are milkfish and shrimps. The reason why we wanted to visit them was that they refuse to use any of the chemical agents or drug and it triggered our curiosity, how do they make it?"),a.default.createElement("div",{className:"innerpart"},"By actually visiting them on the ground, we discovered that they do not use any chemical medicine or drug to kill bacteria, instead, they use Bacillus subtilis to compete with Vibrio. We surprisingly found out this kind of ecological competence is similar to what our wet lab is trying to mimic. Though adding good germs can’t guarantee no fish will be infected, they believed doing so was far better than adding artificial chemicals which have huge uncertainty to humankind. When asking about whether they would detect the concentration of Vibrio, they told us the current method they use to detect Vibrio is coating the plate, which is not efficient and not so accurate."),a.default.createElement("div",{className:"innerpart"},"Furthermore, they provided us with pieces of advice and feedback regarding our Automatic Color-Detecting Examiner (ACE):"),a.default.createElement("div",{className:"innerpart"},a.default.createElement("div",null,"1.ACE needs to be equipped with high accuracy, shutdown notification, and self-calibration."),a.default.createElement("div",null,"2. If they knew the concentration of Vibrio earlier, they can take actions to prevent the Vibrio from growing too much beforehand, such as adding Bacillus subtilis."),a.default.createElement("div",null,"3. Since the growing time of Vibrio to be pathogenic is way longer than one day, a period of sampling doesn’t need to be as frequent as 1 hour, but one day. "),a.default.createElement("div",null,"4. They suggested us to target indoor breeding customers, since that if the device is set outdoors, it may be easily broken down. Moreover, fish with high economic value bred indoor might be more fragile than the outdoors.")),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.interview.eco_2})),a.default.createElement("div",null,"After considering their suggestions, we decided to extend the sampling period to 12 hours, adding the fool-proof design, such as that if the camera shut down or cannot be connected, it will return a “no camera” sign to our application.")))),p=a.default.createElement("div",null,a.default.createElement("div",{className:"part"},a.default.createElement("div",null,a.default.createElement("div",{className:"part-title"},a.default.createElement("a",{href:"http://dataweb.nmmba.gov.tw/ResearchMemberDetail.aspx?CondID=bfcdc17d-da5f-48f8-9af8-819a3376101a"},"Dr. Jimmy Kuo")),"Dr. Jimmy Guo, a vice researcher of the national museum of marine biology and aquarium, have done many types of research of Vibrio in the aquarium. He first shared with us how he made the research on Vibrio and explained why the growth curve of Vibrio was so strange. The reason is that Vibrio is a special species which is different from other normal bacteria. Vibrio will be dormant and make its metabolism disordered when stored at low temperature. Therefore, if we want to get the expectative results, we should culture fresh Vibrio rather than take one from the refrigerator."),a.default.createElement("div",null,"Besides the advice for the wet lab, Dr. Guo also gave us some suggestions to our hardware. He recommended that we could use the battery as our electric power source and thus, avoiding the loss of results from the blackout. Additionally, our device should calibrate regularly, or the results would be deviant. Finally, since our device could detect frequently, we could target on the customers cultivating remunerative aquatic agriculture."),a.default.createElement("div",null,a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.interview.aqua_1})))),a.default.createElement("div",{className:"part"},a.default.createElement("div",null,a.default.createElement("div",{className:"part-title"},a.default.createElement("a",{href:"https://www.nmmba.gov.tw/research/RMenber_Content.aspx?n=291A05EEDDA1D4C4&sms=CF7FFA69EB91FF61&s=6A6B57F5FE966020"},"Dr. Ming-An Tsai ")),"Dr. Ming-An Tsai, an assistant researcher in the national museum of marine biology and aquarium, is interested in doing research on diseases of aquatic animals. Fortunately, Dr. Tsai’s lab is also doing some Vibrio-detecting experiment, so we first exchanged some ideas about Vibrio experiments. After introducing our project, Dr. Tsai gave us some important advice. First of all, he suggested us that we should ensure which way we would use to avoid bacterial diseases, killing Vibrio or inhibiting its growth. Additionally, he reminded us that we should not kill all the Vibrio in the water, or we might destroy the ecological balance. Finally, he gave us a crucial reference that if the concentration of Vibrio in the water goes beyond 104 CFU/mL, it would be more infectious. This data is a vital reference not only in our wet lab but also in our dry lab."),a.default.createElement("div",null,a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.interview.aqua_2}))))),h={viewbar:{Title:"Interview",Subtitle:"Visited the research institute, fishing farm, and aquarium",image:o.default.image.topfull.interview},content:{Introduction:"Our project is an interdisciplinary work which is so comprehensive and complete to makes it possesses a potent potential to practice in the real world. Based on this advantage, to navigate our project toward a suitable direction to either meet the true demand and solve real problem is quite essential. At the beginning, we visited Deputy director of Fisheries Research Institute. He gave us some advice on how to start our project. Then, we had conducted interviews with fishermen in Taiwan, visited the fishing farm with them and received countless useful information about how to alter our design to fit their demand and improve the latest solution they adopt to combat bacterial infection. Moreover, with the advice given by researchers from national museum of marine biology and aquarium, we gained much more practical perspective on what’s the pros and cons of our solution and how we can lower the risk that will harm the environment. With their help, we have achieved the core value of human practice: know who need, find what they need and meet their need.        ",parts:[{Title:"Fisheries Research Institute, COA ",Description:u},{Title:"Interview with Fish Farmer",Description:c},{Title:"Ecological Breeding",Description:d},{Title:"Traditional Fishermen",Description:f},{Title:"National Museum of Marine Biology and Aquarium",Description:p}]}},m=function(e){function t(){return function(e,t){if(!(e instanceof t))throw new TypeError("Cannot call a class as a function")}(this,t),function(e,t){if(!e)throw new ReferenceError("this hasn't been initialised - super() hasn't been called");return!t||"object"!=typeof t&&"function"!=typeof t?e:t}(this,(t.__proto__||Object.getPrototypeOf(t)).apply(this,arguments))}return function(e,t){if("function"!=typeof t&&null!==t)throw new TypeError("Super expression must either be null or a function, not "+typeof t);e.prototype=Object.create(t&&t.prototype,{constructor:{value:e,enumerable:!1,writable:!0,configurable:!0}}),t&&(Object.setPrototypeOf?Object.setPrototypeOf(e,t):e.__proto__=t)}(t,i.Component),r(t,[{key:"render",value:function(){return a.default.createElement(l.default,h)}}]),t}();t.default=m},function(e,t,n){"use strict";Object.defineProperty(t,"__esModule",{value:!0}),t.default=void 0;var r=function(){function e(e,t){for(var n=0;n<t.length;n++){var r=t[n];r.enumerable=r.enumerable||!1,r.configurable=!0,"value"in r&&(r.writable=!0),Object.defineProperty(e,r.key,r)}}return function(t,n,r){return n&&e(t.prototype,n),r&&e(t,r),t}}(),i=n(0),a=s(i),o=(s(n(10)),s(n(4))),l=s(n(8));function s(e){return e&&e.__esModule?e:{default:e}}var u=a.default.createElement("div",null,a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.public.nctu})),a.default.createElement("div",null,"Engaging with other iGEM teams to exchange ideas and to brainstorm together is a crucial part of iGEM. Therefore, we joined a half-day meet-up with three iGEM teams in Taiwan, NTHU_Formosa, NCTU_Formosa, and Mingdao. During the meet-up, we presented our projects, had some discussions with other teams, and received some problems and feedbacks on our projects. After brainstorming, we knew what the problem was and what we could do to improve the project. This meet-up truly broadened our horizons and we got loads out of it. Following are some feedbacks we received from the meet-up:"),a.default.createElement("div",null,a.default.createElement("div",{className:"part"}," 1. There were some drawbacks of our project of biosafety if we put our engineered E. coli into the open water area. After discussion, we finally decided to replace this way by putting it into a close water and to use it with a filter. By doing so, we could effectively control our engineered bacteria and thus, reducing the problem about biosafety. "),a.default.createElement("div",{className:"part"}," 2. We should ensure our device can really benefit the users. Therefore, we visited some local fishermen in person and sought for their suggestions. To find out more, check the ",a.default.createElement("a",{href:"https://2018.igem.org/Team:NTHU_Taiwan/Interview"},"interview")," page"))),c=a.default.createElement("div",null,a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.public.meetup_group})),a.default.createElement("div",null,"In late July, we participated in the first Eurasian meet-up held by BGI college and TU Delft in Shenzhen, China. There are 20 stunning teams from 5 different countries visited Shenzhen, discussing and sharing all kinds of iGEM issues and projects during the conference. Also, we presented and conveyed our idea to other teams, which lets us receive several feedbacks and they were quite useful for our project. At this point, we did find some inspiring ideas which sharpen our current work. Here we conclude something that influenced our work:"),a.default.createElement("div",null,a.default.createElement("div",{className:"part"},"1. The other iGEMers provided us with some useful feedbacks, for instance, they suggested us to integrate and double-check the consistency of data from our bio-reactor and Vibrio-detecting system. Also, team members from CUHK gave us some suggestions about our image detection system. Moreover, some people suggested us other switch systems for our experimental design like toehold switch. After the conference, we searched for several switch systems but we still stick to our original idea because we thought that the disadvantage for our STAR system is the least."),a.default.createElement("div",{className:"part"},"2. A more concise presentation should be considered. This is the first presentation after our frame was formed out, but we spent much time explaining the detailed information about our project. Furthermore, we didn’t integrate our wet lab works and dry lab works in our presentation which was not convincing to the audience. After the presentation, we then modified the whole slides into a more concise and more comprehensible version which do help more audiences to get into our story."),a.default.createElement("div",{className:"part"},"3. The meet-up official set up several workshops and lectures during the conference. For instance, they invited former iGEM judge and iGEM ambassador to share their experiences like “how to be an iGEMer?” from many different perspectives. It seems to us that we learned more details and tips about how to handle the iGEM project and figured out the overview of this competition.")),a.default.createElement("div",null,"Last but not least, except for the feedback we received, we also build a strong friendship connection there. The meet-up official assigns a buddy team for each team and our buddy is Marburg university from Germany. We really enjoyed the moment with them. "),a.default.createElement("div",null,a.default.createElement("div",{className:"gallery"},a.default.createElement("img",{style:{width:"45%",margin:"0px 1%"},src:o.default.image.public.meetup_self}),a.default.createElement("img",{style:{width:"45%",margin:"0px 1%"},src:o.default.image.public.meetup_presentation})))),f=a.default.createElement("div",null,a.default.createElement("div",{className:"illustration"},a.default.createElement("iframe",{style:{height:"500px",width:"100%"},src:"https://www.youtube.com/embed/1wib7JQEZ78"})),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"About TRANS"),a.default.createElement("div",null,"TRANS Conference is a platform where entrepreneurs, experts, investors, and students can exchange ideas, which was created by H. Spectrum, a startup incubator/accelerator focusing on the healthcare sector. As we had a chance to join in this forum, we decided to present our project to people in the healthcare field as many as possible, hoping to get some feedback from them and improve our project."),a.default.createElement("div",null,a.default.createElement("div",{className:"gallery"},a.default.createElement("img",{className:"illust-image",src:o.default.image.public.trans})))),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"part-title"},"Discussion and feedback"),a.default.createElement("div",null,"During TRANS, we found that people who are interested in our project are from multiple different fields and not just professionals. We’ve met students, bacterial professionals, fishery companies and some previous iGEM teams in Taiwan, whose backgrounds are medical, biology, microbiology, culture studies, medical engineering, and electrical engineering."),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.public.discuss_1})),a.default.createElement("div",{className:"part"},a.default.createElement("div",null,"After the discussion, we sorted out the most common questions from our listeners as below:"),a.default.createElement("div",null,a.default.createElement("div",null,"1. What is the principle of MV-kit and what’s its difference between PCR analysis?"),a.default.createElement("div",null,"2. Biosafety such as being eaten by people and genetic mutation of E.coli."),a.default.createElement("div",null,"3. Will engineered genes exchange with bacterial organisms in the environment, causing genetic recombination problems because of the complex environment?"),a.default.createElement("div",null,"4. E.coli residual problems."),a.default.createElement("div",null,"5. Is engineered E.coli regulation acceptable? GMO products in Taiwan are still very opposed, how to persuade?"),a.default.createElement("div",null,"6. The growth and decline effect of E. coli will still have resistance problems in the future. Is there any solution?"))),a.default.createElement("div",{className:"part"},a.default.createElement("div",null,"And we also got some feedback and advice:"),a.default.createElement("div",null,a.default.createElement("div",null,"1. The mechanism of STAR/anti-STAR can also be applied to the sterilization of intensive care units."),a.default.createElement("div",null,"2. We could transform our target to a smaller range to simplify our problem, such as moving our environment from outdoor to indoor."),a.default.createElement("div",null,"3. In the future, if our growth and decline mechanism success, we could experiment our genes by intelligent shrimp culturing."),a.default.createElement("div",null,"4. To solve the problem of drug resistance, try adding multiple attack lines to E. coli."),a.default.createElement("div",null,"5. A company called ID Water which major in water quality testing suggested us to cooperate with them and tried to start an undertaking."))),a.default.createElement("div",{className:"gallery"},a.default.createElement("img",{style:{width:"30%",margin:"0px 1%"},src:o.default.image.public.discuss_2_1}),a.default.createElement("img",{style:{width:"30%",margin:"0px 1%"},src:o.default.image.public.discuss_2_2}),a.default.createElement("img",{style:{width:"30%",margin:"0px 1%"},src:o.default.image.public.discuss_2_3})),a.default.createElement("div",{className:"part"},a.default.createElement("div",{className:"innerpart"},"After digesting the advice, we decided to modify our project in detail by meeting some of the professional companies, which we have saved the contacting information. First, we emailed to ID Water, asking for a chance to visit their company, hoping to have a chance to communicate some detecting methods. For example, we have trouble in analyzing with fluorescence in a bioreactor, which is part of their technique in water analyzing. ID Water thus told us that their cooperator, IGC-BIO, a healthcare company in Taiwan, is also researching in the method of interaction of bacteria, so we also plan to visit them someday."),a.default.createElement("div",{className:"innerpart"},"We could also meet Hin-Chung Wong professor in Soochow University, who is a profession in Vibrio field, for more detailed information about Vibrio to perfect our project. And we have checked again that our project doesn’t violate the iGEM rule. In the future, if we want to test our bacteria, LiveStrong Biomedical Technology promised us that they would provide a shrimp culturing sink for us to practice our experiment. Finally, thank for those who gave us lots of precious advice."),a.default.createElement("div",{className:"gallery"},a.default.createElement("img",{style:{width:"45%",margin:"0px 1%"},src:o.default.image.public.discuss_3_1}),a.default.createElement("img",{style:{width:"45%",margin:"0px 1%"},src:o.default.image.public.discuss_3_2}))))),d=a.default.createElement("div",null,a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.public.xgem})),a.default.createElement("div",null,"Interacting with people from different backgrounds is amazing and we do cherish every moment we engaged with other iGEM teams to exchange ideas as well as share techniques. For us, bringing the people across the world together to brainstorm about synthetic biology and how to make this world a better place is the magic of iGEM. Therefore, we established a platform on Slack named XGEM. On XGEM, teams can share information and ask questions immediately. So far, 18 teams from 14 different countries have joined XGEM. More about the teams joining XGEM:"),a.default.createElement("div",null,a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.public.chart_1})),a.default.createElement("div",{className:"illustration"},a.default.createElement("img",{className:"illust-image",src:o.default.image.public.chart_2})),a.default.createElement("div",{className:"illustration",style:{width:"70%"}},a.default.createElement("img",{className:"illust-image",src:o.default.image.public.chart_3}))),a.default.createElement("div",null,"Its really exciting to have 19 teams from all over the world to interact on XGEM. Some teams have posted their questionnaires related to their project or shared what they had done. We really appreciate your participation and can’t wait to meet these teams in person at Giant Jamboree!")),p={viewbar:{Title:"Public",Subtitle:"Meetups, conferences, and online platform",image:o.default.image.topfull.public},content:{Introducation:"Due to the fact that the synthetic biology is a brand-new field for the general public. Consequently, creating an enlarging division of knowledge between the general public and synthetic biology is necessary, and thus the public engagement is crucial for bridging the gap. The following are the activities we attended this year. We learned so much from the public, our friends and other iGEM teams. 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