<p>Created the first prototype for the team page</p>

<p>Social media created</p>

<p>Social media buttons same color and fixed iGEM style issues</p>

<p>Multiple week summary styles created for the notes</p>

<p>Added the header with mobile compatibility als resized logos and readded them to the header</p>

<p>Added template header and footer with 'under construction' content</p>

<p>Small fixes and menu adjustments</p>

<p>Added a PI list with the secondary PI</p>

<p>Designed and added content holders to the index Page</p>

<p>Added myself to the teammember Page and did some bugfixing</p>

<p>Started working on a better looking menubar</p>

<p>Added the ability to add sub-menus to items in the menu bar. Integration into the website and added indication for which Page you are on</p>

<p>Updated all Pages to the new style with an under construction sign</p>

<p>Added a way to recognise Pages under construction using the menubar</p>

<p>Updated the content containers so that they are responsive</p>

<p>The first draft of the architecture design for the temperature controller was made.</p>

<p>Content added to the human practices page</p>

<p>Content created and added to the main page</p>

<p>Today a few changes have been made to the architecture design of the temperature controller. The choice was made to use a premade power supply as opposed to a self-made one, because this was considering the time left to finish this device the best choice. A few alterations were made to make the architecture design better readable.</p>

<p>A 3d drawing was made for the casing that will hold the temperature controller. The first prototype was printed, but was determent to be inadequate due to the lack of enough space for the cooler block.</p>

<p>Added the remaining instructors to the team</p>

<p>In the drawing space was added to fit the cooler block for the temperature controller. Slits were added to the drawing for the nuts to slide into. The altered drawing was printed. The newly printed parts, cooler block, fan and Peltier element were assembled. Everything fit together.</p>

<p>To start testing the production of CooA, we firstly resuspended and transformed the BioBricks from the kit witch we wanted to use (Table 1). This has been done using the iGEM Kit plate instructions (Protocol 1, step 1 - 3) and the NEB 10 be&#769;ta transformation protocol adapted New England Biolabs (Protocol 2), using bought cells with a reported transformation efficienty of 1-3 10^9 cfu/&mu;g pUC19. Inucbation was at 37&deg;C.<br>

<p>A short test run was done with the newly assembled cooler. The test concluded that the temperature controller could cool a water droplet to the point of freezing and heat it back up to its boiling point.</p>

<p><p>After making a 20 mL overnight culture of our NEB 10 be&#769;ta cells containing our BioBricks, we performed a mini prep (Protocol 3). Using our nanodrop spectrophotometer&nbsp;&nbsp;which gave us the following DNA concentrations:<br>

Also from the previously mentioned overnight culture 1 mL has been used to make Glycerol Stock (Protocol 4)</p>

<p>The choice to use a computer PSU as a power supply was made. The choice was made on the grounds, that computer PSU have a standard 12, 5 and 3.3V output. This is beneficial because these are the voltages necessary for the selected components.</p>

<p><p>Today we performed a digestion, dephosphorilation and ligation (following protocols 4 and 5) of a few promotor and RBS BioBricks. Because of administrative compliations only the variants with J23113 and a RBS could be made.</p>

<p>Changes were made to the architecture design of the temperature controller. Different voltage regulators were selected to fit the newly chosen power supply. To better readability of the architecture design a legend was added.</p>

<p>Custom footprints were made for multiple components and added to the parts library in the PCB design software.</p>

<p>The provisional electronic circuit design for the temperature controller was made.</p>

<p>The electronic circuit design for the temperature controller was finished. Measuring point were added to the electronic circuit design.</p>

<p>To have some basic biobricks ready, we stocked them by doing a plasmid isolation and freeze the DNA. </p>

<p>To start on a basic construct we did a digestion and ligation with the parts we isolated earlier. To prevent original constructs we did a defosphorylation after the digestion. We controlled the digestion by doing a gelelktroforeses. </p>

<p>Started working on the board layout for the PCB of the temperature controller.</p>

<p>We continued with the ligated DNA by transformating it to Neb10Beta. This was plated onto agarplates and incubated for 1 day. </p>

<p>Further work was done to the board layout for the temperature controller.</p>

<p>The results of 7/12/2018 were collected, in action of this we decided to plate some ligated and transferred DNA from 7/12/2018 again. This we did because there wasn't a clear grow of colonies on the plates.</p>

<p>We tried to let bacteria grow on paper, therefore we used our own buisnesscards. We let the buisnesscards absorb some LB-medium and put them inside petridishes. </p>

<p>Work has been done on the board layout for the temperature controller. The board layout is now finished.</p>

<p>To know if the ligation was succesfull we will controll the DNA, therefor we ented the bacteria's with DNA in LB-medium to grow overnight. We also plated the used bacteria's onto new plates to use for further experiments. </p>

<p>We did a blue white screening on our buisnesscards to see if bacteria had grew on them. We transferred the buisnesscards to new petrydishes and added ITPG and X-gal to the petrydishes. </p>

<p>Frontend design for the notebook</p>

<p>The resulst of 7/16/2018 showed that the DNA didnt ligate as planned.  To be able to digest enough DNA we did a plasmid isolation again. We digested this DNA and froze it for further use. </p>

<p>We did another round of plasmid isolation for stocking. </p>

<p>Designed a way to follow experiments that take up multiple days and eventually designed a notebook entry page and made it a responsive page</p>

<p>We digested all DNA we have in stock to test for our basic construct. We tested the DNA by doing a gelelktroforeses. </p>

<p>Created a one page notebook. Made the notebook entry page more suited for the entry length</p>

<p><p>Transformations of the&nbsp;biobricks K133071, K173003 and I13453 are performed. K133071 will produce CO2 if there's pyruvate present and K173003 will produce CO2 if urea is present. I13453 is a promotor which will work if there's arabinose present.&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp;&nbsp;<br>

<p class="experiment"><p><span style="font-size: 14px;"><strong>Experiment 1: Transformation biobricks</strong></span><br>

Transformation of the biobricks K133071, K173003 and I13453 in NEB10B�ta.<br>

- Heater (42�C) &nbsp;<br>

- Incubator + shaker (37�C and 250 rpm)&nbsp;<br>

<p>To be able to use the DNA from the biobricks, they were first diluted in Nuclease free water. This was done by adding 10ul of nuclease free water to the DNA and incubated for 5 minutes at room temperature (18-24�C). After incubation the DNA is transferred to Eppendorf tubes&nbsp;and put on ice.&nbsp;<br>

For the transformation 1 ul of the DNA was added&nbsp;to 50 ul&nbsp;chemically competent cells strain&nbsp;NEB10Beta. The mixture was incubated for 30 minutes on ice. The mixture got a heat shock by putting the tube&nbsp;in a heater (42�C) for 30 seconds. After the heat shock the tube was immediately put on ice for 5 minutes.&nbsp;<br>

For good grow of the cells, 950 ul of outgrow medium was added to the mixture. After mixing the cells with outgrow medium, the culture was incubated for 1 hour at 37�C and 250 rpm.&nbsp;&nbsp;<br>

After incubation the culture was plated onto LB-agar plates. For specific growth of bacteria there was antibiotics added to the plates. The antibiotics that were used, depended on the plasmid. The biobricks used in this experiment were resistent to chloramphenicol (Can). We worked with a work concentration of 35 ug/ml for chloramphenicol. For good results we plated 100 ul undiluted culture onto a plate and centrifuged the rest of the culture. The supernatant was discarded and the pellet was resuspended and plated onto a LB-agar plate. The plates were then incubated by 37�C.&nbsp;<br>

The second transformation from the biobricks was successful. The bacteria with the biobricks plasmids will be stored by 4�C and can be used for further experiments.&nbsp;<br>

<p>New biobricks, used to make an ATP sensor, were transformed into NEB10Beta. The culture was than plated onto agar and incubated for 1 day.  </p>

<p>New biobricks, which produces gasses, were transformed into NEB10Beta. The culture was than plated onto agar and incubated for 1 day.  </p>

<p>Started working on an automatic notebook creator</p>

<h1 class="title">Transformation</h1>

<p><p>Today we did a transformation with&nbsp;new biobricks. The biobricks were built into chemically competent cells from the strain NEb10Beta. After transformation the culture was plated on agar plates with&nbsp;antibiotics. The results were checked the next day.&nbsp;</p>

<p class="experiment">Materials:&nbsp;

- Heater (42�C) &nbsp;<br>

- Incubator + shaker (37�C and 250 rpm)&nbsp;<br>

To be able to use the DNA from the biobricks, they were first diluted in Nuclease free water. This was done by adding 10ul of nuclease free water to the DNA and incubated for 5 minutes at room temperature (18-24�C). After incubation the DNA is transferred to Eppendorf tubes&nbsp;and put on ice.&nbsp;<br>

For the transformation 1 ul of the DNA was added&nbsp;to 50 ul&nbsp;chemically competent cells strain&nbsp;NEB10Beta. The mixture was incubated for 30 minutes on ice. The mixture got a heat shock by putting the tube&nbsp;in a heater (42�C) for 30 seconds. After the heat shock the tube was immediately put on ice for 5 minutes.&nbsp;<br>

For good grow of the cells, 950 ul of outgrow medium was added to the mixture. After mixing the cells with outgrow medium, the culture was incubated for 1 hour at 37�C and 250 rpm.&nbsp;&nbsp;<br>

After incubation the culture was plated onto LB-agar plates. For specific growth of bacteria there was antibiotics added to the plates. The antibiotics that were used, depended on the plasmid. The biobricks used in this experiment were resistent to chloramphenicol (Can). We worked with a work concentration of 35 ug/ml for chloramphenicol. For good results we plated 100 ul undiluted culture onto a plate and centrifuged the rest of the culture. The supernatant was discarded and the pellet was resuspended and plated onto a LB-agar plate. The plates were then incubated by 37�C.&nbsp;<br>

The plates are stored by 4�C.&nbsp;<br>

The transformation from the biobricks was successful. The bacteria with the biobricks plasmids will be stored by 4�C and can be used for further experiments.&nbsp;<br>

<p>Automatic generation of entries completed</p>

<p>Used the Google drive REST API to download the entries from drive</p>

<p>Used the igemwiki API to update pages using software</p>

<h1 class="title">Transformation</h1>

<div class="attendees">Loraine Nelson | Elise Grootscholten | Paul Reusink</div>

<p><p>To be able to always get the necessary biobricks, we made an glycerol stock after every transformation. The glycerol stocks are stored at -80 degrees Celsius. If needed, they can retrieved from this storage to use for experiments.&nbsp;</p>

</p>

<p class="experiment"><p>Making glycerol stocks of the biobricks.&nbsp;<br>

Materials:&nbsp;<br>

- 87% glycerol&nbsp;<br>

- culture with biobrick plasmids<br>

- LB-medium<br>

- Variable Volume Pipette<br>

- Sterile pipetpoints<br>

Method:&nbsp;<br>

The colonies from the transformation were used to make an overnight culture. The culture was made by putting a colony in LB-medium and then the culture was&nbsp;incubated for a night at 37�C. &nbsp;<br>

The next day the cultures had grown close and were ready for a glycerol stock. For making glycerol stocks there was added 250 ul 87% glycerol to 1 ml culture. This was frozen in -80�C. &nbsp;<br>

<br>

The stocks were stored at -80�C in drawer 3, tower 1, drawer&nbsp;</p>

</p>

<div class="number"><b>02</b></div>

<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Transformation</h3>

<h1 class="title">Assessing Gas Production BioBricks in E.Coli</h1>

<div class="attendees">Elise Grootscholten | Randall de Waard</div>

<p><p>For further experiments there is&nbsp;isolated DNA needed of the biobricks J23100, K133071, K173003 and I134353. The DNA is isolated out of the bacteria with the help of a plasmid purifaction kit. After isolation this DNA can be used for digestion and ligation or other experiments.&nbsp;<span style="font-size:12px;">J23100 (from the glycerolstock): 325,59 ng/ul, K173003: 217,06 ng/ul, K133071: 186,79 ng/ul,&nbsp;</span>I13453: 88,18 ng/ul</p>

<p class="experiment"><p><span style="font-size:14px;"><strong>Experiment 3: Minipreps</strong></span><br>

<span style="font-size:12px;">Minipreps of the following biobricks in NEB10B�ta:</span><br>

-J23100: Constitutive promoter<br>

-K133071: Urea --&gt; ammonia + CO2<br>

-K173003: Pyruvate --&gt; acetaldehyde + CO2<br>

-I134353: Promoter (AraC protein binds with arabinose)<br>

<span style="font-size:14px;"><strong>Method:&nbsp;</strong></span><br>

After the transformation we made an overnight culture from the colonies. The colonies were anted into 20 ml LB and incubated overnight at&nbsp;37�C and 150 rpm. The culture was mini prepped the next day. For isolation, 5,4 ml was added step wise in a 2ml Eppendorf cup. The culture was then centrifuged at &gt;12000x g for 30 seconds. The supernatant was deposed, and the pellet was resupended with A1 buffer and vortexed. To the resuspended culture was <strong><span style="font-size:14px;">then.....???<br>

Results</span></strong><br>

<span style="font-size:12px;">J23100: 325,59 ng/ul<br>

K173003: 217,06 ng/ul<br>

K133071: 186,79 ng/ul</span><br>

I13453: 88,18 ng/ul</p>

<div class="number"><b>03</b></div>

<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Assessing Gas Production BioBricks in E.Coli</h3>

<p>Connected the page uploader to the notebook generator</p>

<p>Created a program that uploads all updated pages</p>

<p>Built a page builder that automatically adds the header and footer before uploading</p>

<p>Redone the google API handler for reusability and readability and made the google API handler generic</p>

<p>revamped the main program</p>

<p>Finished redoing the application for generating and uploading pages</p>

<p>Added month categorization to the page</p>

<p><p>For further experiments we needed isolated DNA. The DNA is isolated out of the bacteria with the help of a plasmid purifaction kit. After isolation this DNA can be used for digestion and ligation or other experiments.</p>

<p class="experiment"><p>Materials:&nbsp;<br>

After the transformation we made an overnight culture from the colonies. The colonies were anted into 20 ml LB and incubated overnight at&nbsp;<span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">37�C and 150 rpm. The culture was mini prepped the next day. For isolation, 5,4 ml was added step wise in a 2ml Eppendorf cup. The culture was then centrifuged at &gt;12000x g for 30 seconds. The supernatant was deposed, and the pellet was resupended with A1 buffer and vortexed. To the resuspended culture was then.&nbsp;</span></p>

<p><p>The biobricks&nbsp;J23100, K133071, K173003 and I134353 were&nbsp;sucessfully digested after the second time. After the&nbsp;digestions the&nbsp;biobricks K133071 and K173003&nbsp;were dephosphorylated and ligated with the inserts J23100 and I13453. This was done in the original backbone of K133071 and K173003, and not another control backbone. To know&nbsp;if the biobricks were right ligated this was done by testing practically.&nbsp;See the experiments:&nbsp;<em>Testing gas production.</em>&nbsp;</p>

<p class="experiment"><p><span style="font-size: 14px;"><strong>Experiment 4: Digestion, Defosforylation and&nbsp;ligation</strong></span><br>

<p><p><span style="font-size:14px;">Testing different amounts of urea and sodiumpyruvate to know which concentrations the bacteria survive.</span></p>

<p class="experiment"><p><span style="font-size: 16px;"><strong>Experiment 1:&nbsp;Testing different concentrations of Urea and Sodium pyruvate</strong></span><br>

<p><p>Transformations of the biobricks (K133071 + J23100), (K13071 + I1345), (K173003 + J23100) and (K173003 + I13453) in NEB10B�ta.&nbsp;There was no grow except for the biobrick combination K133071 + J23100.</p>

<p class="experiment"><p><span style="font-size: 16px;"><strong>Experiment 5: Transformation</strong></span></p>

-Chemo competent cells NEB10b�ta (New England Biolabs)</span></span><br>

<p><p>A colony PCR is done for the NEB10B�ta E.coli cells with expected&nbsp;the biobrick combination of K133071 with J23100. Nevertheless, on a gel the difference with and without promotor couldn't be seen. So there must be another way of proving the right biobricks are there.</p>

<p class="experiment"><p><span style="font-size: 14px;"><strong>Expermiment 6: Colony PCR K133071 + J23100</strong></span><br>

<p>The ligations and transformations of the biobricks K173003 + J23100, K173003 + I13453, and K133071 + I13453 have been performed again.&nbsp;There was a lot of grow on the agar plates. To know&nbsp;if the biobricks were right ligated this was done by testing practically.&nbsp;See the experiments:&nbsp;<em>Testing gas production.</em>&nbsp;</p>

<p class="experiment"><p><span style="font-size: 14px;"><strong>Experiment 7: Ligation of biobricks</strong></span><br>

<p><p>Making a set up for the gas production testing and testing it with NEB10B�ta with the expected biobricks in it (K133071 + J23100) and a negative control.</p>

<p class="experiment"><p><span style="font-size: 14px;"><strong>Experiment 1: Gasproduction testing</strong></span><br>

Making a set up for the gas production testing and testing it with NEB10B�ta with the expected biobricks in it (K133071 + J23100) and a negative control.<br>

Making a set up for the gas production testing and testing it with NEB10B�ta with the expected biobricks in it (K133071 + J23100).<br>

<p><p>Making a set up for the gas production testing and testing it with NEB10B�ta with the expected biobricks in it (K133071 + I13453), (K173003 + J23100), (K173003 + I13453)&nbsp;and a negative control.</p>

<p class="experiment"><p><span style="font-size: 14px;"><strong>Experiment 2: Gasproduction testing </strong></span><br>

Making a set up for the gas production testing and testing it with NEB10B�ta with the expected biobricks in it (K133071 + I13453), (K173003 + J23100), (K173003 + I13453)&nbsp;and a negative control<br>

<p><p>Gasproduction testing for the biobricks (K173003 + I13453), (K133071 + I13453) and negative controls (B0015 and K133071 without urea and arabinose).<br>

<p class="experiment"><p><span style="font-size:14px;"><strong>Experiment 3: Gasproduction testing</strong></span><br>

<p><p>Testing the gasproduction&nbsp;of the colonies&nbsp;9 and 10 of biobricks (K173003 + J23100), colonies 5 and 7 of biobricks (K173003 + I13453) and colony 4 of&nbsp;biobricks (K133071 + I13453) with and without centrifuging the bacteria. There is also a negative control&nbsp;(J04450 pSB1K3) with Kanamycine. The negative contol started with a lot of gas inside the tube. We can not see wether there is produced more after a day or not. This have to be tested later.</p>

<p class="experiment"><p><span style="font-size:14px;"><strong>Experiment 4: Gasproduction testing&nbsp;</strong></span><br>

<p><p>Minipreps are made of colonies 11 and 19 of biobrick combination&nbsp;K133071 + J23100.&nbsp;Results: 11. 270,57 ng/ul&nbsp;19. 253,59 ng/ul.</p>

<p class="experiment"><p><span style="font-size:14px;"><strong>Minipreps</strong></span><br>

<p><p>A miniprep of the biobrick K352002 is made. The concentration is 69,77 ng/ul.</p>

<p class="experiment"><p><strong>Experiment 9: Minipreps</strong><br>

<p><p><span style="font-size:14px;">Testing different amounts of urea and sodiumpyruvate to know which concentrations the bacteria survive.</span></p>

<p class="experiment"><p><span style="font-size: 16px;"><strong>Experiment 2:&nbsp;</strong></span><strong><span style="font-size:16px;">Testing different concentrations of Urea and Sodium pyruvate</span></strong><br>

&nbsp;</p>

<p><p>Digestions, gelelectrophoresis, dephosphorylations and ligations of different biobrick combinations. These biobrick combinations are 4 different promoters (J23100, I13453, K352002, K352003)&nbsp; with 4 different gasproduction biobricks (k173003, K173013, K133071, K133116). J23100 is about 1 kb to long. The rest seems likely to be right digested. The ligations will be transformed in NEB10b�ta and digested again as control.</p>

<p class="experiment"><p><span style="font-size:14px;"><strong>Experiment 10: Digestion, defosforylation and ligation</strong></span><br>

&nbsp;</p>

<p><p>Transformations of the biobrick&nbsp;combinations in NEB10b�ta. The transformations are plated on kanamycine agar plates, because all the ligations were done in pSB1K3 (kanamycine resistence) backbone. Pink and with colonies appeared after incubation by 37 degrees Celcius for about 12h. The white colonies will be used for further experiments.</p>

<p class="experiment"><p><strong>Experiment 11: transformation of the biobrickombinations in NEB10b�ta.</strong><br>

<p><p>Plasmid DNA was isolated of 20 different colonies. The first time something went wrong. The second time we had good concentrations of isolated plasmid DNA. After this we will digest the DNA to see if the plasmids all have the required biobricks. The tube have a code from now on, see table 1.<br>

<p class="experiment"><p><strong>Experiment&nbsp;12: Overnight cultures and minipreps</strong><br>

&nbsp;</p>

<p><p>The minipreps of experiment 12 are digested with the restriction enzymes: SmaI and ScaI. Only Sca and Sma are both incubated at 37 degrees Celcius. Sma has to be incubated at 25 degrees Celcius. This is done in experiment 15.</p>

<p class="experiment"><p><strong>Experiment 12: Digestions of the minipreps</strong><br>

Therefore, Sma was added again and was incubated at 25 degrees Celcius. See results experiment 16.</p>

<p><p>Digestions have been done of the biobrickcombinations&nbsp;K352002 +K133116 + pSB1K3 and K352003 + K133116 + pSB1K3 with restriction-enzymes ScaI and SmaI</p>

<p class="experiment"><p><strong>Experiment 14: Digestions</strong><br>

<p><p>All the digestions of experiment 13 have digested again with SmaI for an our at 25 degrees Celcius. Because earlier the digestions have been put immediately bt 37 degrees Celcius. It looks like H1, J1 and D2 include the right biobricks.&nbsp;</p>

<p class="experiment"><p><span style="font-size:14px;"><strong>Experiment 16: Gelelectroforesis</strong></span><br>

<p><p>Minipreps of the biobrick combinations:&nbsp;K352002+K133116+pSB1K3 (1) and&nbsp;K352003+K133116+pSB1K3 (2) are done. The DNA concentration is for 1:189,3 ng/&mu;l and for 2: 355,3&nbsp;ng/&mu;l.</p>

<p class="experiment"><p><span style="font-size:14px;"><strong>Experiment 14: Minipreps</strong></span><br>

<p><p>Different&nbsp;strains&nbsp;of E.coli are tested as negative control. Those strains were:&nbsp;NEB10B�ta, BL21 (DE3), BL21, HB101, DH5alpha and&nbsp;JM109. NEB10B�ta produced the most gas. BL21, HB101 and JM101 produced none/almost none gas. Because BL21 isn't a K12 strain we can not use that one. That is why we will test further gasproduction in HB101 and JM109. And maybe those will be our final E.coli strains.</p>

<p class="experiment"><p><span style="font-size:14px;"><strong>Experiment 5: Testing different strains&nbsp;of E.coli</strong></span><br>

Make overnight cultures of the following E.coli tribes: NEB10B�ta, BL21 (DE3), BL21, HB101, DH5alpha, JM109.<br>

<p>32 minipreps have been performed. A lot of them have a low concentration nucleic acid and/or have a high 260/280 and 260/230 rate. Therefore those will be done again.</p>

<p class="experiment"><span style="font-size:14px;"><strong>Experiment 17: Minipreps</strong></span><br>

Thise are: A1, A4, A5, A6, B1, B5, B6 C1, C2, C3, C4, C5, C6, E1, E2, E5, E6, F4, F5</p>

<p><p>Making overnight cultures of the numbers: A1, A4, A5, A6, B1, B5, B6 C1, C2, C3, C4, C5, C6, E1, E2, E5, E6, F4, F5 with Kanamycin. These biobrick combinations can be found in experiment 12. Also&nbsp;the first digestions with XbaI, Eco0109I, HindIII and SspI-HF have been done.</p>

<p class="experiment"><span style="font-size:14px;"><strong>Experiment 19: Making overnight cultures + first digestions</strong></span><br>