Difference between revisions of "Team:Edinburgh OG/Human Practices"

Latest revision as of 03:57, 18 October 2018

PhagED: a molecular toolkit to re-sensitise ESKAPE pathogens

Integrated Human Practices

Over the course of summer, The University of Edinburgh OG iGEM 2018 team had learned about the importance of plastic in our lives, making it virtually impossible to detach it completely due to its versatile attributes and application. However, we also learned that plastic waste is detrimental to our environment. To tackle this problem, we chose to focus our efforts on bioplastic production, specifically the copolymer PHBV. As a group, we firmly believe that to approach the plastic issue it is necessary to design environmentally responsible products. For that reason, it was essential to engage in conversation with experts in the field, from bioplastic consultants to plastic producers. As a result of the discussions with various stakeholders, the OG iGEM team designed genetically modified parts and integrated a sustainability analysis to Valeris.Ed. Thus, for our Integrated Human Practices we wanted to show how the dialogue with multiple sources shaped our project and how it evolved from the initial idea to the final project we are presenting. Following this, in the left-side section, we summarised how stakeholders’ interaction lead to changes in the design of the project. On the other hand, in the right section and organised in chronological order, we listed the consulted stakeholders and the valuable insights they provided to the project.

The team has not only focused on how to contribute in solving the plastic problem, but it also has thought of the potential use of Synthetic Biology in other fields. Accordingly, we collaborated with other iGEM teams and organised different presentations to promote Synthetic Biology and the iGEM competition in our local and international community, i.e. young students in Macau. We are happy to have spread the wonder of this world and work with the future first iGEM team in Macau.

Dialogue with

During the last lecture for the course Tools of Synthetic Biology the team explained to our classmates about the iGEM competition through an introduction of the iGEM history and principles. We also presented our idea of producing bioplastics from local materials with the purpose of delivering a sustainable process.

Feedback

Our classmates pointed out the importance of bioplastics and challenged us of to understand how synthetic biology could improve a process that has been done using biotechnology. They also suggested to look into the whisky industry or wastewater as potential raw materials.

Dr Lorraine Kerr - Commercial Relations Manager at School of Biological Sciences, University of Edinburgh

Dr. Kerr was our principal contact between stakeholders and experts in the bioplastic field. During meetings we discussed our project, objectives and approach to work out the story and how to talk to stakeholders. She advised us to use the expert's experience and point of view to get more specific information, define targets and to set goals prior to interviewing.

Dr Adrian Higson - Director and Lead Consultant for Biobased Products, NNFFCC The Bioeconomy Consultants

Plastics are one of the most versatile materials that are immensely beneficial to our society (e.g. food packaging which helps in reducing food wastage). However, their life cycle constitutes a big challenge. Current proposals to confront this issue include recycling, the reduction of plastic consumption, and a change from plastic to bioplastic. Nonetheless, it is necessary to consider the type of bioplastic and its life cycle. On top that, it is not recommended to consider bioplastics as carbon sink, instead we should look into the circular economy where plastic is use multiple times instead of one, in addition to the design of products where recycling could be possible. The major barrier in bioplastic production is the economy of scale – the increase in innovation can only happen with the increase in production, however the current condition does not permit that due to low sales.

Assessing the sustainability

Dr Adrian Higson discussed the necessity of plastics and the problems of their current life cycle. We expressed our major concerns; the feedstock materials and the sustainability of the process. As a result, the project gained great insight from the experience of Dr. Higson. He commented that in terms of feedstock, the use of agriculture materials is simple not feasible. The use of side products however is more sustainable and by using them we are making a good choice of raw material. We need to look at the energy efficiency, water efficiency, and conversion process. The GHG emissions and water footprint are not the only impacts to look at, we have to consider the social and economic sustainability of the production.

We also discussed the possible end-of-life scenarios, according to him the recycling of bioplastics and the final break of the polymer into monomers constitute a splendid way of tackling the waste problem. This also increases the chain value of the bioplastic as the monomers can be sold as chemicals for another industry.

Implementation

Dr Adrian Higson supported our idea of using by-products from other industry (whisky to be exact) due to its sustainability features. However, he pointed the necessity of looking in detail to some other impacts. This conversation was followed by a team discussion, where, with the advice of our supervisors we started to research the incorporation of the Life Cycle Assessment model to look into the life cycle of our bioplastic.

After of our talks with experts and stakeholders we incorporated the LCA tool to measure the impacts of each aspect of our product life.

Life Cycle Assessment is a systematic way of quantifying the environmental sustainability of a product and helps to identify aspects for improvement. It is a vital tool for sustainability studies and if implemented correctly can act as the basis to evaluate different scenarios and facilitate decisions.

Implementations

The LCA resulted in an iterative tool that began as the repercussion of the dialogues with experts and ended as a decision making instrument. The LCA supported the idea of using the in-situ secretion system and showed us that the recycling scenario modelled for our process contributed to decrease the environmental impacts up to 70%. To read more about our LCA results please click here.

Dr. James Hallinan, Dr. Steve Thomas & Dr. James Brown,– Experts in Synthetic Biology and Material Sciences, Cambridge Consultants

The interviewed experts endorsed our project and explained that PHAs represent an innovating example of the delivery of sustainable products through cutting edge technology. They suggested the group to focus on the properties, design and potential applications of PHAs instead of tackling the production price. Furthermore, they emphasised the manufacturing challenges of producing PHAs, which are the major hurdle preventing their wide scale adoption: the scalability.

Assessing Feedstock and Downstream Processing

It was also discussed the topic relating to the present obstacles in bioplastic production, according to them feedstock resources and downstream processing are the key problems that must be addressed immediately. Feedstock is an environmentally sustainable way to sequester carbon source. Additionally, the interviews added the attractiveness of using underutilised products from industries as raw materials, because they are more budget friendly since they are usually under-priced, however, it needs to be built on an effective logistics strategy in terms of the usage as well as the consistency of the components in the by-product itself. The downstream processing is one of the biggest bottlenecks in the process, for this reason having a method for delivering the bioplastic without the necessity of disrupting the cells would increase the efficiency of the overall production and potentially can lead to the implementation of continuous-batch fermentation which could run for more hours and the carbon source and the product can be simultaneously added and removed, respectively.

Implementation and Impacts in our project

First, the use of whisky by-products as raw material is supported by the consistency of their elements. Second, the logistics of using them can be more achievable as they are the product of a local industry. The in-situ secretion system constitutes a potential method for reducing the use of hazardous chemicals in the process hence reduces the environmental impact of the PHBV production. Moreover, the potential use of continuous-batch fermentation can contribute to the overall efficiency of the manufacture. The cost of the bioplastic production is not the most important factor, due to the necessity of shift from oil to bio-based raw material. We took their advice of looking more into the properties and we decided to only focus on PHBV production because PHBV is a PHA copolymer with a larger thermal processing window, thus less brittle than PHB.

We were invited to the  iGEM Northern Meet-up organised by the St Andrews iGEM team. This event was intended for the cooperation between teams as a space to share ideas, experiences and feedback for the projects. During the event we got feedback from professors and previews generations of iGEMers.

Implementation

They gave us advice of how to construct our narrative and to shape our objectives. Regarding the design of the process, they suggested to look into the use of fed-batch or continuous fermentation in order to increase the yield of PHBV. Furthermore, they also suggested alternatives for our in silico model.

Morag Garden - Head of Sustainability & Innovation, Scotch Whisky Association

Morag facilitated the group a lot of information about the local situation of the whisky market and their by-products. She highlighted the increasing global demand for Scotch and the now 128 distillers operating across Scotland's five different whisky regions. In addition, she gave us suggestions for alternative community members looking at valorising the distillery materials, as they would be a useful source of information. Additionally, she also mentioned their interest in the outcomes of the research.

Some notes from the Whisky by-products study case

The whisky sector remains high with a growth rate estimated at 2.1% over the period to 2018. This would result in an approximate increase in by-products in the region of: Draff – 52,800 tonnes and Pot ale – 88,800 tonnes. By-products from Scotch Whisky distillery (draff and pot ale for example), have long been reused by the agriculture sector as a valuable animal feed and fertiliser for agricultural land (Figure 1). However, suitable routes would be need to be found for these by-products, as existing are near their natural capacity. New developments have recently opened up new markets for the use of by-products as feedstock for renewable heat or its use to make protein feed for salmon farming and biofuel.

Figure 1 Availability of by-product feedstocks by local authorities in Scotland. Taken from Ricardo Energy & Environment for Zero Waste Scotland (2017). Biorefining Potential for Scotland.

Figure 2. The current uses for whisky by-products in Scotland. Taken from Zero Waste Scotland (2015). Sector Study on Beer, Whisky and Fish. Final Report

A circular economy approach is to use resources more efficiently, keep resources in use for as long a time as possible and to minimise waste. Identifying opportunities to improve circular approaches, for example keeping by-product materials within Scotland for further processing. There are many potential products that could feasibly be made from by-products, adding further value to a business.

References

Miller, R. The landscape for biopolymers in packaging. Miller- Klein Associates report Summary and Full Report available from The National Non-Food Crops Centre, Heslington, New York, UK, 2005, nnfcc.co.uk.
Scottish Environment Protection Agency (2018). Scotch Whisky Sector Plan. Available at: https://consultation.sepa.org.uk/communications/sector-approach-to-regulation-consultations-on-sco/supporting_documents/SEPA_Whisky%20Sector%20Plan_Final.pdf
Ricardo Energy & Environment for Zero Waste Scotland (2017). Biorefining Potential for Scotland. Available at: https://www.zerowastescotland.org.uk/sites/default/files/Biorefining%20Potential%20for%20Scotland%20Final%20report.pdf

Through this visit, we learnt about the whisky production and collected whisky production by-products samples (i.e. pot ale and draff) to be used as carbon source in our experiments.

We were introduced to the whisky production through the tour throughout the distillery. We learnt that the whisky industry managed their resources locally and sustainably and ensured that minimum waste is generated in every batch of the production. The information obtained was very helpful in conducting the Life Cycle Assessment.

Figure 1 (from top to bottom) Tour guide explained about the kiln. Samples of malt before separated into husk, grist, and flour. Our team collected samples.

Dr. Guo-Qiang Chen - Director of Center for Synthetic and Systems Biology, Tsinghua University and Chief Scientist at BLUEPHA

Jin Yin - Technique Manager at BLUEPHA

They endorsed our project explaining that our work is conducive to the long-term development of the world. As global environmental awareness increases, the market for biodegradable plastics will certainly develop. This includes the PHA market and research is needed to achieve this shift from plastics to bioplastics, especially in Europe where composting action is encouraged by the government. PHAs certainly has this unique advantage over other bioplastics.

Over the interviews with Jin Yin and Dr. Chen, it was discussed the ways of solving the plastic pollution. In their point of view, one of them is promoting the use of bio-based plastics for containers and bags. However, they emphasised the necessity of research institutes and industry to work in the optimisation of the manufacturing processes and strain properties. For them, the bottlenecks in the PHA production are the strain properties, as the ability of using different raw materials, and the downstream process which is limiting the efficiency and purity of the product. As SynBio practitioners, they believe that the answer is in the use of this discipline to help solving these problems.

Bluepha Co., Ltd. is a synthetic biology and biomaterial company established in Beijing, China. The company was the first SynBio Startup Founded by iGEMers, and they were the winner of the Future Planet Award for Sustainable Growth. Moreover, they are pioneering a revolutionary cost-effective method for producing bioplastics.

Jin Yin and Dr. Chen:

“Over 20 companies from different countries are focusing on the optimisation of PHA production, all of us are trying hard on solving many practical problems and we truly believe that the answer of some of these problems is in the use of Synthetic Biology. However, it is also important to focus on the discovery of medical applications of PHAs. The biocompatibility of PHAs enclose the potential for using it as artificial cartilage and nerve conducts. For example, US has permit the usage of P(4HB) in surgery. They believe that PHAs is a new thing deserving further studies with surprising properties that can be a potential answer for diseases such like osteoporosis. Despite of the current drawbacks, we believe in a more optimistic future for PHAs“

Implementations

We confirmed that our in-situ secretion system is a good approach to tackle the downstream processing problem. As further directions, they recommended us to look for improvements to the strain for being able to use whisky by-products more efficiently. However, it is important to note, that solving technical issues of PHA production is as important as it is related to the biomedical research. As a result, a new focus for our future research is to look into new biomedical applications of PHAs and specifically PHBV.

Paul Tan – Officer SINOPEC Guangzhou, China

Sinopec Guangzhou Petrochemical Company (“Guangzhou Company” for short) is one of the leading petrochemical enterprises in South China. The core business of Guangzhou Company covers refining products including solid plastics, such as polyethylene, polypropylene and polystyrene.

Jianfeng and Paul endorsed our project explaining that they welcome the technology development of bioplastic, even as a competitor to them. The mass convenience of plastic in food, clothes, car industry, etc. are too important and so far there is no other material that can replace plastic. And, currently the degradation of plastic is still too expensive for its price and its environmental impact. As a result, bioplastics constitute a possible solution to the growing demand of plastics.

Understanding the recycling market of plastics

Jianfeng and Paul have experience in the petroleum-based industry and knowledge of how the plastic market has change. We were aiming to understand the importance of different disposable scenarios and they gave us an insight of the recycling industry. They explained that after the reduction of oil prices in 2014, the recycling industry had retreated as making new plastic became much cheaper than recycling it. In addition, they mentioned that recycled plastics are slightly different and the manufacturers need to add other materials to improve the physical properties. These additives tend not be in the labels of the plastics making the recycling process more difficult in a further stage.

Implementations

The recycling scenario would not be the best end-of-life for our plastics, in conjugation with the comments of Dr. Higson from NNFCC we believe that a combination of recycling and composting could be helpful in terms of the sustainability of our bioplastics. The results of the research in PHBV degradation and composability should be incorporated to the design of PHBV products.

Paul Mines– CEO at Biome Technologies

“Plastics: wonder materials or existential threat”

Paul Mines began the lecture at The University of St Andrews by disclosing the issue of plastic pollution and the ubiquity of plastic due to its cheap and useful properties. Plastics have been around since 70 years ago, replacing other materials and expanding the plastic market. Then he continued discussing how bioplastic products can help to solve plastic pollution by offering plant-based and biodegradable alternatives to petroleum-derived plastics.

Understanding the impacts of the process

Regarding the carbon sequestration in bioplastics, the net carbon footprint is less and bioplastic can function as CO₂ storage, but however, it cannot help with the GHG problem in the world. Furthermore, he talked about the possible disposal scenarios such as composting, anaerobic digestion and incineration. Finally, he ended his presentation by emphasising on what needs to be developed to achieve the wide use of bioplastic. Some of his examples were: Plant science, biomass production, pre-treatment of the biomass, monomer treatment, and tests and scale up. Nevertheless, the market cannot go forward if the impact of bioplastics products to the environment and society is not understood.

The lecture reaffierded that ‘Human Practices’ aspect is crucial in every project design and not just iGEM. Through our Human Practices journey, we found LCA tool which assessed the environmental impacts of a product and/or process, which is in line with what Paul Mines mentioned.

Difference between revisions of "Team:Edinburgh OG/Human Practices"

Latest revision as of 03:57, 18 October 2018

Integrated Human Practices

Dialogue with

Dialogue with

Dialogue with

Dialogue with

MSc students in Tools of Synthetic Biology class
2017/2018

Commercial Links

The National Non-Food Crops Centre (NNFCC)

Life Cycle Assessment (LCA)

Cambridge Consultants

iGEM Northern UK Meet-up @ U St Andrews

Scotch Whisky Association

Visit to Local Whisky Distillery in Edinburgh

BLUEPHA Technologies

SINOPEC

Biome Technologies

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-<h1 style="text-align: center;"><strong>Notebook</strong></h1>
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-<h4 style=" font-size:150% " "><strong>Investigation of the best gene order of <em>phaCAB</em>&nbsp; for PHB production(Pepper)</strong></h4>
+<h1 style="text-align: center;"><strong>Integrated Human Practices</strong></h1>
+<div class="column two_third_size">
+<p style="text-align: justify;">Over the course of summer, The University of Edinburgh OG iGEM 2018 team had learned about the importance of plastic in our lives, making it virtually impossible to detach it completely due to its versatile attributes and application. However, we also learned that plastic waste is detrimental to our environment. To tackle this problem, we chose to focus our efforts on bioplastic production, specifically the copolymer PHBV. As a group, we firmly believe that to approach  the plastic issue it is necessary to design environmentally responsible products. For that reason, it was essential to engage in conversation with experts in the field, from bioplastic consultants to plastic producers. As a result of the discussions with various stakeholders, the OG iGEM team designed genetically modified parts and integrated a sustainability analysis to <em>Valeris.Ed</em>. Thus, for our Integrated Human Practices we wanted to show how the dialogue with multiple sources shaped our project and how it evolved from the initial idea to the final project we are presenting. Following this, in the left-side section, we summarised how stakeholders’ interaction lead to changes in the design of the project. On the other hand, in the right section and organised in chronological order, we listed the consulted stakeholders and the valuable insights they provided to the project.</p>
-   <div class="container"><button class="btn btn-primary" type="button" data-toggle="collapse" data-target="#tittle11">
+<p style="text-align: justify;">The team has not only focused on how to contribute in solving the plastic problem, but it also has thought of the potential use of Synthetic Biology in other fields. Accordingly, we collaborated with other iGEM teams and organised different presentations to promote Synthetic Biology and the iGEM competition in our local and international community, i.e. young students in Macau. We are happy to have spread the wonder of this world and work with the future first iGEM team in Macau.&nbsp;&nbsp;</p>
-<p><strong>Week 1- Week 2 Primers design and experimental preparation</strong></p></button>
-        <div id="tittle11" class="collapse">
-<ul>
+<div class="col-sm-4">
-<li>Design and order primers</li>
-<p>Primers for changing gene order in <em>pha </em>operon are designed in Gibson Assembly website</p>
-<li>Preparation of M9 medium</li>
-<li>Primers stock solution preparation</li>
-</ul>
+<div class="btn-group text-align">
+<p style="text-align: center;"><img style="display: block; margin-left: auto; margin-right: auto;" src="https://static.igem.org/mediawiki/2018/9/9c/T--Edinburgh_OG--IHP_-_rev1half.png" width="650" height="280" /></p>
+<h4 style="text-align: center; font-size: 200%;"><strong>Dialogue with</strong></h4>
+<p style="text-align: center;"><button class="btn btn-info" type="button" data-toggle="collapse" data-target="#tittle3">Tools</button> &nbsp; <button class="btn btn-info" type="button" data-toggle="collapse" data-target="#tittle1">NNFCC</button> &nbsp; <button class="btn btn-info" type="button" data-toggle="collapse" data-target="#tittle6">Cambridge Consultant</button>&nbsp; <button class="btn btn-info" type="button" data-toggle="collapse" data-target="#tittle8">Scotch Whisky Assc</button>&nbsp; <button class="btn btn-info" type="button" data-toggle="collapse" data-target="#tittle2">BluePHA</button></p>
+<p style="text-align: center;"><img style="display: block; margin-left: auto; margin-right: auto;" src="https://static.igem.org/mediawiki/2018/d/d6/T--Edinburgh_OG--IHP_-_rev1bottom.png" width="650" height="185" /></p>
+<p style="text-align: center;">&nbsp;</p>
+&nbsp; &nbsp; &nbsp;<br />
+<div class="btn-group text-align" style="text-align: center;">
+<p><img style="display: block; margin-left: auto; margin-right: auto;" src="https://static.igem.org/mediawiki/2018/5/5c/T--Edinburgh_OG--IHP_-_rev2top.png" width="650" height="280" /></p>
+<h4 style="text-align: center; font-size: 200%;"><strong>Dialogue with</strong></h4>
+<p style="text-align: center;"><button class="btn btn-info" type="button" data-toggle="collapse" data-target="#tittle6">Cambridge Consultant</button> &nbsp; <button class="btn btn-info" type="button" data-toggle="collapse" data-target="#tittle1">NNFCC</button> &nbsp; <button class="btn btn-info" type="button" data-toggle="collapse" data-target="#tittle7">Northern UK Meetup</button></p>
+<p style="text-align: center;"><img style="display: block; margin-left: auto; margin-right: auto;" src="https://static.igem.org/mediawiki/2018/9/90/T--Edinburgh_OG--IHP_-_rev2bottom.png" alt="" width="650" height="385" /></p>
+<p>&nbsp;</p>
+<div class="btn-group text-align">
+<p><img style="display: block; margin-left: auto; margin-right: auto;" src="https://static.igem.org/mediawiki/2018/1/18/T--Edinburgh_OG--IHP_-_rev3top.png" width="650" height="280" /></p>
+<h4 style="text-align: center; font-size: 200%;"><strong>Dialogue with</strong></h4>
+<p style="text-align: center;"><button class="btn btn-info" type="button" data-toggle="collapse" data-target="#tittle5">LCA</button> &nbsp; <button class="btn btn-info" type="button" data-toggle="collapse" data-target="#tittle6">Cambridge Consultant</button> &nbsp; <button class="btn btn-info" type="button" data-toggle="collapse" data-target="#tittle2">BluePHA</button></p>
+<p><img src="https://static.igem.org/mediawiki/2018/8/80/T--Edinburgh_OG--IHP_-_rev3bottom.png" width="650" height="246" /></p>
+<div class="btn-group text-align">
+<p>&nbsp;</p>
+<p><img src="https://static.igem.org/mediawiki/2018/b/bf/T--Edinburgh_OG--IHP_-_rev4top.png" width="650" height="164" /</p>
+<h4 style="text-align: center; font-size: 200%;"><strong>Dialogue with</strong></h4>
+<p style="text-align: center;"><button class="btn btn-info" type="button" data-toggle="collapse" data-target="#tittle5">LCA</button> &nbsp; <button class="btn btn-info" type="button" data-toggle="collapse" data-target="#tittle2">BluePHA</button> &nbsp; <button class="btn btn-info" type="button" data-toggle="collapse" data-target="#tittle11">Biome Tech</button> &nbsp; <button class="btn btn-info" type="button" data-toggle="collapse" data-target="#tittle10">Sinopec </button>&nbsp;</p>
+<p><img src="https://static.igem.org/mediawiki/2018/7/74/T--Edinburgh_OG--IHP_-_rev4bottom.png" width="650" height="435" /></p>
+<p>&nbsp;</p>
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-   <div class="container"><button class="btn btn-primary" type="button" data-toggle="collapse" data-target="#tittle12">
-<p><strong>Week 3- Week 5 Building and confirming constructs</strong></p></button>
-        <div id="tittle12" class="collapse">
-<ul>
-<li>Amplify and extract the fragments of pSB1C3 backbone, pSB1C3-phaB</li>
-<li>Gel electrophoresis and Gel extraction</li>
-<li>Amplify and extract fragments of <em>phaA</em>, <em>phaB</em>, <em>phaC</em> with different overlap (failed)</li>
-<p>Due to the special primer design from Gibson, except backbone fragment, all of the other fragments were amplified by primers with overhang. In this case, a problem was caused by wrong identity of annealing sequence and overlap sequence. Therefore, it is necessary to produce fragments without overlap firstly, and used them as template, in order to avoid the impact of the overlap and annealing sequence.</p>
-<li>Design and order new primers for <em>phaA</em>, <em>phaB</em> and <em>phaC</em> gene template without overhang</li>
-<li>Purification of pSB1C3 backbone and pSB1C3-phaB DNA fragments</li>
-<li>Amplify <em>phaA</em>, <em>phaB</em> and <em>phaC</em> gene templates by PCR followed by gel electrophoresis and gel extraction</li>
-</ul>
 </div>
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+    </div>
+    <div class="col-sm-4">
+    </div>
+    <div class="col-sm-4">
+<!--1. MSc students @ Tools -->
-   <div class="container"><button class="btn btn-primary" type="button" data-toggle="collapse" data-target="#tittle13">
-<p><strong>Week 6- Week 8</strong></p></button>
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-<ul>
-<li>Amplify separative <em>phaA</em>, <em>phaB</em>, and <em>phaC</em> genes fragments with different overhangs</li>
-<li>Gel electrophoresis of <em>phaA</em>, <em>phaB</em>, and <em>phaC</em> genes fragments</li>
-<li>Assemble fragments to build new constructs using Gibson Assembly and transformation (<em>&nbsp;E. coli</em> DH5&alpha; competent cells)</li>
-<li>colony PCR and double digestion</li>
-</ul>
-<p><img style="display: block; margin-left: auto; margin-right: auto;" src="https://static.igem.org/mediawiki/2018/c/c8/T--Edinburgh_OG--Notebook_-_Pep1a.png" width="584" height="215" /><strong><br /><img style="display: block; margin-left: auto; margin-right: auto;" src="https://static.igem.org/mediawiki/2018/b/b0/T--Edinburgh_OG--Notebook_-_Pep1b.png" width="487" height="338" /></strong></p>
-<p style="text-align: left;"><strong>Figure 1. Results of colony PCR. </strong>The fragments of 4kb size were expected to be the positive results (a)BCA are 12 colonies from plates expressed by <em>phaBAC</em> plasmid. (b)BCA are 12 colonies from <em>phaBCA </em>plasmid. (c) represents 12 colonies from <em>phaABC</em> expression plasmid. 11 colonies in (d) are from <em>phaCBA</em> plasmid. (e) is the result if negative control of this PCR experiment, which represents colonies from expression of empty vector. (f) shows 8 colonies from<em> phaACB</em> plasmid. Finally, (g) is the second negative control of purity water used in this reaction, no band means dH<sub>2</sub>O used is pure.</p>
-<ul start="5">
-<li>Confirmation new plasmids by double digestion. Transfer new plasmids into another strain E. coli BL21(DE3)</li>
-</ul>
+    <div class="container"><button class="btn btn-warning" type="button" data-toggle="collapse" data-target="#tittle3" aria-expanded="false" aria-controls="tittle3"><h3><strong> &#32; &nbsp;MSc students in Tools of Synthetic Biology class <br> 2017/2018   </strong></h3></button>
+        <div id="tittle3" class="collapse">
+   <div class="col-sm-5">
+<blockquote>
+<p style="text-align: justify;">During the last lecture for the course Tools of Synthetic&nbsp;Biology the&nbsp;team explained to our classmates about the&nbsp;iGEM&nbsp;competition&nbsp;through an introduction of the&nbsp;iGEM&nbsp;history and principles. We also presented&nbsp;our idea of producing bioplastics from local materials with the purpose of delivering a sustainable process.</p>
+     </blockquote>
+<div class="textbox">&nbsp;</div>
+<div class="container"><button class="btn btn-info" type="button" data-toggle="collapse" data-target="#MSc">Read more</button>
+<div id="MSc" class="collapse">
+  <div class="col-sm-5">
+<p><strong>Feedback</strong></p>
+<p style="text-align: justify;">Our classmates pointed out the importance of bioplastics and challenged us of to understand how synthetic biology could improve a process that has been done using biotechnology.&nbsp;They also suggested to look into&nbsp;the whisky industry or wastewater as potential raw materials.&nbsp;</p>
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-<p><strong>Week 9- Week 10 PHB production and extraction </strong></p></button>
-        <div id="tittle14" class="collapse">
-<ul>
-<li>Culture the <em> coli</em> strain harboured <em>phaACB</em> operon or <em>phaCBA</em> operon to produce PHB (250 ml flasks)</li>
-<li>The optical density was measured every 3 hours for plotting growth curves. Spread bacteria that harboured new constructs <em>phaBAC</em>, <em>phaBCA</em>, <em>phaCBA</em>, <em>phaACB</em>, <em>phaABC</em> or original<em> phaCAB</em> operon on Nile Red plate. Culture E. coli that contained new constructs of <em>phaBAC,</em> <em>phaBCA,</em> <em>phaCBA, phaACB</em> or <em>phaABC</em> for Nile Red plate reader assay.</li>
-<li>Optical density measurement and fluorescent intensity measurement</li>
-<li>Extract produced PHB from <em> coli</em> that harboured <em>phaACB</em> operon or <em>phaCBA </em>operon after 72h cultivation</li
-<p><strong><img style="display: block; margin-left: auto; margin-right: auto;" src="https://static.igem.org/mediawiki/2018/d/dc/T--Edinburgh_OG--Notebook_-_Pep2.png" width="353" height="213" /></strong></p>
-<p style="text-align: center;"><strong>Figure 2 Nile red plates images of E. coli that harboured new constructs. </strong>On each plate, half plate is spread by negative control- pSB1C3 empty vector, which should not have any fluorescence, another half is plated with new operon plasmids culture.</p>
-<p>&nbsp;</p>
-<p style="text-align: center;"><strong>Table 1 </strong>OD value detected by Nile Red plate reader</p>
-<table style="margin-left: auto; margin-right: auto;" width="0">
-<tbody>
-<tr>
-<td style="text-align: center;" width="58">
-<p><strong>Hours</strong></p>
-</td>
-<td style="text-align: center;" width="97">&nbsp;</td>
-<td style="text-align: center;" width="87">
-<p><strong>BAC</strong></p>
-</td>
-<td style="text-align: center;" width="86">
-<p><strong>BCA</strong></p>
-</td>
-<td style="text-align: center;" width="90">
-<p><strong>CBA</strong></p>
-</td>
-<td style="text-align: center;" width="102">
-<p><strong>ACB</strong></p>
-</td>
-<td style="text-align: center;" width="95">
-<p><strong>ABC</strong></p>
-</td>
-</tr>
-<tr>
-<td style="text-align: center;" width="58">
-<p><strong>24 h</strong></p>
-</td>
-<td style="text-align: center;" width="97">
-<p>2% Glucose</p>
-</td>
-<td style="text-align: center;" width="87">
-<p>0.45&plusmn;0.02</p>
-</td>
-<td style="text-align: center;" width="86">
-<p>0.48&plusmn;0.01</p>
-</td>
-<td style="text-align: center;" width="90">
-<p>0.65&plusmn;0.01</p>
-</td>
-<td style="text-align: center;" width="102">
-<p>0.69&plusmn;0.01</p>
-</td>
-<td style="text-align: center;" width="95">
-<p>0.42&plusmn;0.01</p>
-</td>
-</tr>
-<tr>
-<td style="text-align: center;" width="58">&nbsp;</td>
-<td style="text-align: center;" width="97">
-<p>3% Glucose</p>
-</td>
-<td style="text-align: center;" width="87">
-<p>0.42&plusmn;0.01</p>
-</td>
-<td style="text-align: center;" width="86">
-<p>0.44&plusmn;0.01</p>
-</td>
-<td style="text-align: center;" width="90">
-<p>0.64&plusmn;0.01</p>
-</td>
-<td style="text-align: center;" width="102">
-<p>0.63&plusmn;0.01</p>
-</td>
-<td style="text-align: center;" width="95">
-<p>0.47&plusmn;0.01&nbsp;</p>
-</td>
-</tr>
-<tr>
-<td style="text-align: center;" width="58">
-<p><strong>45 h</strong></p>
-</td>
-<td style="text-align: center;" width="97">
-<p>2% Glucose</p>
-</td>
-<td style="text-align: center;" width="87">
-<p>0.66&plusmn;0.01</p>
-</td>
-<td style="text-align: center;" width="86">
-<p>0.70&plusmn;0.01</p>
-</td>
-<td style="text-align: center;" width="90">
-<p>0.94&plusmn;0.02</p>
-</td>
-<td style="text-align: center;" width="102">
-<p>0.99&plusmn;0.02</p>
-</td>
-<td style="text-align: center;" width="95">
-<p>0.61&plusmn;0.01&nbsp;</p>
-</td>
-</tr>
-<tr>
-<td style="text-align: center;" width="58">&nbsp;</td>
-<td style="text-align: center;" width="97">
-<p>3% Glucose</p>
-</td>
-<td style="text-align: center;" width="87">
-<p>0.66&plusmn;0.01</p>
-</td>
-<td style="text-align: center;" width="86">
-<p>0.68&plusmn;0.02</p>
-</td>
-<td style="text-align: center;" width="90">
-<p>0.94&plusmn;0.01</p>
-</td>
-<td style="text-align: center;" width="102">
-<p>0.98&plusmn;0.03</p>
-</td>
-<td style="text-align: center;" width="95">
-<p>0.68&plusmn;0.01&nbsp;</p>
-</td>
-</tr>
-</tbody>
-</table>
-<p style="text-align: justify;">Table 1 is the OD600 value of Nile Red plate reader, culture fed with 2% glucose and 3% glucose are incubated for 24h and 45h, then stained by Nile Red, and placed into a black 96 well microplate and read by Tecan Infinate M200 (provided by Edinburgh Genome Foundry). Excitation wavelength is 520 nm, and emission wavelength is 590 nm. Data are average number &plusmn; SEM (standard error of mean), n=3. P-value&lt;0.05.<strong>&nbsp;</strong></p>
-<p style="text-align: center;"><strong>Table 2 </strong>Fluorescence intensity measurement by Nile red plate reader</p>
-<table style="margin-left: auto; margin-right: auto;" width="0">
-<tbody>
-<tr>
-<td style="text-align: center;" width="71">
-<p><strong>Hours</strong></p>
-</td>
-<td style="text-align: center;" width="108">&nbsp;</td>
-<td style="text-align: center;" width="85">
-<p><strong>BAC</strong></p>
-</td>
-<td style="text-align: center;" width="85">
-<p><strong>BCA</strong></p>
-</td>
-<td style="text-align: center;" width="95">
-<p><strong>CBA</strong></p>
-</td>
-<td style="text-align: center;" width="94">
-<p><strong>ACB</strong></p>
-</td>
-<td style="text-align: center;" width="104">
-<p><strong>ABC</strong></p>
-</td>
-</tr>
-<tr>
-<td style="text-align: center;" width="71">
-<p><strong>24 h</strong></p>
-</td>
-<td style="text-align: center;" width="108">
-<p>2% Glucose</p>
-</td>
-<td style="text-align: center;" width="85">
-<p>1,683&plusmn;132</p>
-</td>
-<td style="text-align: center;" width="85">
-<p>2,371&plusmn;90</p>
-</td>
-<td style="text-align: center;" width="95">
-<p>35,876&plusmn;850</p>
-</td>
-<td style="text-align: center;" width="94">
-<p>41,757&plusmn;586</p>
-</td>
-<td style="text-align: center;" width="104">
-<p>4,534&plusmn;311</p>
-</td>
-</tr>
-<tr>
-<td style="text-align: center;" width="71">&nbsp;</td>
-<td style="text-align: center;" width="108">
-<p>3% Glucose&nbsp;</p>
-</td>
-<td style="text-align: center;" width="85">
-<p>1,758&plusmn;48</p>
-</td>
-<td style="text-align: center;" width="85">
-<p>2,437&plusmn;108</p>
-</td>
-<td style="text-align: center;" width="95">
-<p>40,330&plusmn;134</p>
-</td>
-<td style="text-align: center;" width="94">
-<p>31,486&plusmn;670</p>
-</td>
-<td style="text-align: center;" width="104">
-<p>4,651&plusmn;222</p>
-</td>
-</tr>
-<tr>
-<td style="text-align: center;" width="71">
-<p><strong>45 h</strong></p>
-</td>
-<td style="text-align: center;" width="108">
-<p>2% Glucose</p>
-</td>
-<td style="text-align: center;" width="85">
-<p>5,374&plusmn;97</p>
-</td>
-<td style="text-align: center;" width="85">
-<p>5,432&plusmn;236</p>
-</td>
-<td style="text-align: center;" width="95">
-<p>29,874&plusmn;685</p>
-</td>
-<td style="text-align: center;" width="94">
-<p>&nbsp;42,295&plusmn;793</p>
-</td>
-<td style="text-align: center;" width="104">
-<p>5,054&plusmn;182</p>
-</td>
-</tr>
-<tr>
-<td style="text-align: center;" width="71">&nbsp;</td>
-<td style="text-align: center;" width="108">
-<p>3% Glucose&nbsp;</p>
-</td>
-<td style="text-align: center;" width="85">
-<p>5,415&plusmn;270</p>
-</td>
-<td style="text-align: center;" width="85">
-<p>5,164&plusmn;41</p>
-</td>
-<td style="text-align: center;" width="95">
-<p>27,890&plusmn;989</p>
-</td>
-<td style="text-align: center;" width="94">
-<p>28,311&plusmn;1988</p>
-</td>
-<td style="text-align: center;" width="104">
-<p>4,499&plusmn;95</p>
-</td>
-</tr>
-</tbody>
-</table>
-<p style="text-align: justify;">Table 2 shows the fluorescence value detected by Nile red plate reader, samples stained by Nile Red, and placed into a black 96 well microplate and read by Tecan Infinate M200 (provided by Edinburgh Genome Foundry). Excitation wavelength is 520 nm, and emission wavelength is 590 nm. Data are average number &plusmn; SEM (standard error of mean), n=3, P-value&lt;0.05.</p>
-<p><strong>The Production of PHB</strong></p>
-<p><strong><img style="display: block; margin-left: auto; margin-right: auto;" src="https://static.igem.org/mediawiki/2018/0/04/T--Edinburgh_OG--Notebook_-_Pep3.png" width="398" height="145" /></strong></p>
-<p style="text-align: center;"><strong>Figure 3 </strong>Image of extracted PHB after cultivation of 45 hours<strong>&nbsp;- </strong>Figure 3 indicates the PHB products extracted from <em>E. coli</em> BL21 harboring six pha operons and empty vector plasmid, samples shown in figure 3 were incubated for 45h</p>
-<p>&nbsp;<img style="display: block; margin-left: auto; margin-right: auto;" src="https://static.igem.org/mediawiki/2018/e/e6/T--Edinburgh_OG--Notebook_-_Pep4.png" width="304" height="193" /></p>
-<p style="text-align: center;"><strong>Figure 4 </strong>Image of extracted PHB after cultivation of 72 hours - Figure 4 shows the PHB products from <em>E. coli</em> BL21 harboring phaCBA and phaACB, which incubated for 72h</p>
-<p style="text-align: center;"><strong>Table 3 </strong>Dry weight of produced PHB and melting temperature</p>
-<table style="margin-left: auto; margin-right: auto;" width="0">
-<tbody>
-<tr>
-<td width="180">
-<p style="text-align: center;"><strong>Operon construct </strong></p>
-</td>
-<td style="text-align: center;" width="114">
-<p><strong>Dry weight</strong></p>
-</td>
-<td style="text-align: center;" width="225">
-<p><strong>Melting temperature start</strong></p>
-</td>
-<td width="85">
-<p style="text-align: center;"><strong>Finish</strong></p>
-</td>
-</tr>
-<tr>
-<td colspan="4" width="605">
-<p><em>&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; Yield of PHB after 45h cultivation</em></p>
-</td>
-</tr>
-<tr>
-<td width="180">
-<p>Empty vector</p>
-</td>
-<td style="text-align: center;" width="114">
-<p>~0</p>
-</td>
-<td style="text-align: center;" width="225">
-<p>/</p>
-</td>
-<td style="text-align: center;" width="85">
-<p>/</p>
-</td>
-</tr>
-<tr>
-<td width="180">
-<p><em>phaCAB </em>origin order</p>
-</td>
-<td style="text-align: center;" width="114">
-<p>0.237 g</p>
-</td>
-<td style="text-align: center;" width="225">
-<p>150</p>
-</td>
-<td style="text-align: center;" width="85">
-<p>185</p>
-</td>
-</tr>
-<tr>
-<td width="180">
-<p><em>phaACB</em></p>
-</td>
-<td style="text-align: center;" width="114">
-<p>0.201 g</p>
-</td>
-<td style="text-align: center;" width="225">
-<p>156</p>
-</td>
-<td style="text-align: center;" width="85">
-<p>180</p>
-</td>
-</tr>
-<tr>
-<td width="180">
-<p><em>phaCBA</em></p>
-</td>
-<td style="text-align: center;" width="114">
-<p>0.072 g</p>
-</td>
-<td style="text-align: center;" width="225">
-<p>160</p>
-</td>
-<td style="text-align: center;" width="85">
-<p>186</p>
-</td>
-</tr>
-<tr>
-<td width="180">
-<p><em>phaBCA</em></p>
-</td>
-<td style="text-align: center;" width="114">
-<p>0.021 g</p>
-</td>
-<td style="text-align: center;" width="225">
-<p>158</p>
-</td>
-<td style="text-align: center;" width="85">
-<p>184</p>
-</td>
-</tr>
-<tr>
-<td colspan="4" width="605">
-<p><em>&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; Yield of PHB after 72h cultivation</em></p>
-</td>
-</tr>
-<tr>
-<td width="180">
-<p><em>phaACB</em></p>
-</td>
-<td style="text-align: center;" width="114">
-<p>0.044 g</p>
-</td>
-<td style="text-align: center;" width="225">
-<p>148</p>
-</td>
-<td style="text-align: center;" width="85">
-<p>165</p>
-</td>
-</tr>
-<tr>
-<td width="180">
-<p><em>phaCBA</em></p>
-</td>
-<td style="text-align: center;" width="114">
-<p>0.18 g</p>
-</td>
-<td style="text-align: center;" width="225">
-<p>176</p>
-</td>
-<td style="text-align: center;" width="85">
-<p>180</p>
-</td>
-</tr>
-</tbody>
-</table>
-<p>Table 3 indicates the dry weight and melting temperature of all PHB products extracted in this project</p>
-<p>&nbsp;</p>
-<p>&nbsp;</p>
 </div>
 </div>
+</div>
+</div>
-<h2><strong>Improving the production of PHBV by introducing <em>bktB</em>&nbsp; to <em>E. coli</em> (Ming)</strong></h2>
+<!--2.  Lorraine -->
-   <div class="container"><button class="btn btn-primary" type="button" data-toggle="collapse" data-target="#tittle21">
-<p><strong>Week 1- Week 2 Designing the fragments and primers </strong></p></button>
-        <div id="tittle21" class="collapse">
-<ul>
-<li>A <em>bktB</em> gene from <em> eutropha </em>H16 was codon-optimised for <em>E. coli</em> and synthesised by IDT in two parts (B1 and B2 fragments) with 696 bp overlap.</li>
+    <div class="container"><button class="btn btn-warning" type="button" data-toggle="collapse" data-target="#tittle4" aria-expanded="false" aria-controls="tittle4"><h3><strong> &#32; &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;Commercial Links &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;    </strong></h3></button>
-<li>Order the primers for fragments amplification</li>
+        <div id="tittle4" class="collapse">
-<li>M9 medium and 30% glucose stock solution preparation</li>
-</ul>
+   <div class="col-sm-5">
+<blockquote>
+<p><strong>Dr&nbsp;Lorraine&nbsp;Kerr</strong> -&nbsp;Commercial Relations Manager at School of Biological Sciences, University of Edinburgh</p>
+<p style="text-align: justify;">Dr. Kerr was our principal contact between stakeholders and experts in the bioplastic field. During meetings we discussed our project, objectives and approach to work out the story and how to talk to stakeholders. She advised us to use the expert's experience and point of view to get more specific information, define targets and to set goals prior to interviewing.</p>
+     </blockquote>
+</div>
 </div>
 </div>
@@ Line 528: / Line 168: @@
-   <div class="container"><button class="btn btn-primary" type="button" data-toggle="collapse" data-target="#tittle22">
-<p><strong>Week 3- Week 5 constructs establishment and culture condition optimization</strong></p></button>
-        <div id="tittle22" class="collapse">
-<ul>
-<li>Establish a new constuct: pSB1C3-phaCAB-bktB (cloning strategy was shown below)</li>
-<p><img style="display: block; margin-left: auto; margin-right: auto;" src="https://static.igem.org/mediawiki/2018/1/1c/T--Edinburgh_OG--Notebook_-_bktb_1.png" alt="" width="611" height="153" /></p>
-<p style="text-align: center;"><strong>Figure 1</strong>. B1 and B2 fragments were codon-optimised for <em>E. coli</em> and synthesised by IDT. The 696 bp overlap was in green and restriction enzyme sites were coloured by red. Two pairs of primers were ordered from IDT to amplify <em>bktB</em>, among which B1 forward primer and B2 reverse primer were used to amplify bktB fragments.</p>
-<p style="text-align: center;">&nbsp;</p>
-<li>Make competent cells (<em>&nbsp;E. coli</em> BL21 and <em>E. coli</em> DH5)</li>
+<!--3.  NNFCC  -->
-<li>Determine the tolerance of high concentration of propionic acid.</li>
-<p>Three concentrations (8mM, 32mM and 48mM) were tested using <em>E. coli</em> cells that harboured pSB1C3-phaCAB.</p>
-<li>Determine the suitable glucose concentration for cultivation. (0.5%, 1%, 2%, 3%, 5%, 10%, 20%)</li>
-<li>Establish new control plasmid pSB1C3 by cutting of the RFP gene.</li>
-<p>&nbsp;</p>
-<p><strong><img style="display: block; margin-left: auto; margin-right: auto;" src="https://static.igem.org/mediawiki/2018/d/de/T--Edinburgh_OG--Notebook_-_bktb_2.png" alt="" width="461" height="162" /></strong></p>
-<p style="text-align: center;"><strong>Figure 2</strong>. Plasmid of control plasmid pSB1C3-RFP. The new control plasmid pSB1C3 was obtained by digesting pSB1C3-RFP with restriction enzyme: <em>Not</em>I followed by self-ligation.</p>
-<p>&nbsp;</p>
-<li>Establish another construct pSB1C3-phaCB-bktB</li>
-</ul>
-<p style="text-align: left;">&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;<img src="https://static.igem.org/mediawiki/2018/8/87/T--Edinburgh_OG--Notebook_-_bktb_3_left.png" alt="" width="235" height="234" /><img src="https://static.igem.org/mediawiki/2018/d/df/T--Edinburgh_OG--Notebook_-_bktb_3_right.png" alt="" width="253" height="230" /></p>
-<p style="text-align: center;"><strong>Figure 3</strong>. The <em>bktB</em> fragments and psB1C3-phaCB-bktB fragments were highlighted by red squares.</p>
-<p>&nbsp;</p>
-</div>
-</div>
+<div class="container"><button type="button" class="btn btn-warning"  data-toggle="collapse" data-target="#tittle1">
+<h3><strong>&nbsp;&nbsp;&nbsp; The National Non-Food Crops Centre (NNFCC)&nbsp;&nbsp;&nbsp;&nbsp;</strong></h3>
+      </button>
+        <div id="tittle1" class="collapse">
+   <div class="col-sm-5">
+<blockquote>
+   <p><strong>Dr Adrian Higson</strong> - Director and Lead Consultant for Biobased Products, NNFFCC The Bioeconomy Consultants</p>
+<p class="cd-timeline-content">Plastics are one of the most versatile materials that are immensely beneficial to our society (e.g. food packaging which helps in reducing food wastage). However, their life cycle constitutes a big challenge. Current proposals to confront this issue include recycling, the reduction of plastic consumption, and a change from plastic to bioplastic. Nonetheless, it is necessary to consider the type of bioplastic and its life cycle. On top that, it is not recommended to consider bioplastics as carbon sink, instead we should look into the circular economy where plastic is use multiple times instead of one, in addition to the design of products where recycling could be possible. The major barrier in bioplastic production is the economy of scale &ndash; the increase in innovation can only happen with the increase in production, however the current condition does not permit that due to low sales.</p>
+          </blockquote>
+<div class="container"><button class="btn btn-info" type="button" data-toggle="collapse" data-target="#NNFCC">Read more</button>
+<div id="NNFCC" class="collapse">
+  <div class="col-sm-5">
+<p><strong>Assessing the sustainability</strong></p>
+<p>Dr Adrian Higson discussed the necessity of plastics and the problems of their current life cycle. We expressed our major concerns; the feedstock materials and the sustainability of the process. As a result, the project gained great insight from the experience of Dr. Higson. He commented that in terms of feedstock, the use of agriculture materials is simple not feasible. The use of side products however is more sustainable and by using them we are making a good choice of raw material. We need to look at the energy efficiency, water efficiency, and conversion process. The GHG emissions and water footprint are not the only impacts to look at, we have to consider the social and economic sustainability of the production.</p>
+<p>We also discussed the possible end-of-life scenarios, according to him the recycling of bioplastics and the final break of the polymer into monomers constitute a splendid way of tackling the waste problem. This also increases the chain value of the bioplastic as the monomers can be sold as chemicals for another industry.</p>
+<p><strong>Implementation</strong></p>
+<p>Dr Adrian Higson supported our idea of using by-products from other industry (whisky to be exact) due to its sustainability features. However, he pointed the necessity of looking in detail to some other impacts. This conversation was followed by a team discussion, where, with the advice of our supervisors we started to research the incorporation of the Life Cycle Assessment model to look into the life cycle of our bioplastic.</p>
+    </div>
+         </div>
+  </div>
+    </div>
+          </div>
+         </div>
+<!--4.  LCA -->
-   <div class="container"><button class="btn btn-primary" type="button" data-toggle="collapse" data-target="#tittle23">
-<p><strong>Week 6 &ndash; Week 7.&nbsp; Nile Red quantitative measurement </strong></p></button>
-        <div id="tittle23" class="collapse">
-<ul>
-<li>Transformation of two new constructs.</li>
-<p>The transformation was tested by double digestion and colony PCR</p>
-<li>Order the Nile red fluorescent dye and prepare the stock solution of 1 mg/ml</li>
-<li>Test whether Nile red can be used for PHA production confirmation and establish new Nile red semi-quantitative measurement.</li>
-</ul>
-<p>&nbsp;<img style="display: block; margin-left: auto; margin-right: auto;" src="https://static.igem.org/mediawiki/2018/5/50/T--Edinburgh_OG--Notebook_-_bktb_4.png" alt="" width="489" height="207" /></p>
-<p style="text-align: center;"><strong>Figure 4</strong>. Nile red staining plate was exposed to the blue light and UV light respectively. High fluorescent was observed on the right part (cells containing pSB1C3-phaCAB plasmid) due to the produced PHA.</p>
-<p>&nbsp;</p>
-</div>
-</div>
-   <div class="container"><button class="btn btn-primary" type="button" data-toggle="collapse" data-target="#tittle24">
-<p><strong>Week 8 &ndash; week 9 cell culture and PHA production</strong></p></button>
-        <div id="tittle24" class="collapse">
-<ul>
-<li>Culture the E.<em>coli</em> strain BL21 harbouring pSB1C3-phaCAB-bktB or pSB1C3-PhaCB-bktB in M9 medium that contained 3% glucose, 8mM propionic acid and 10mM acetic acid.</li>
-<li>Optical density of culture was measured at 16 hours, 24 hours. 32 hours, 48 hours and 56 hours.</li>
-<li>Measure fluorescent intensity of cultures to provide real-time information of PHA production. (semi-quantitative Nile red measurement)</li>
-<p>&nbsp;<img style="display: block; margin-left: auto; margin-right: auto;" src="https://static.igem.org/mediawiki/2018/b/b1/T--Edinburgh_OG--Notebook_-_bktb_5.png" alt="" width="769" height="177" /></p>
-<p style="text-align: center;"><strong>Figure 5</strong>. Samples were collected at different cultivation times and stained by Nile red fluorescent dye. The fluorescent intensity increased during the cultivation times.</p>
-<p>&nbsp;</p>
+    <div class="container"><button class="btn btn-warning" type="button" data-toggle="collapse" data-target="#tittle5"><h3><strong> &#32; &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Life Cycle Assessment (LCA)&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; &nbsp;&nbsp;&nbsp;&nbsp;</strong></h3></button>
+        <div id="tittle5" class="collapse">
+   <div class="col-sm-5">
+<blockquote>
+<p style="text-align: justify;">After of our talks with experts and stakeholders we incorporated the LCA tool to measure the impacts of each aspect of our product life.</p>
+     </blockquote>
+<div class="container"><button class="btn btn-info" type="button" data-toggle="collapse" data-target="#LCA">Read more</button>
+<div id="LCA" class="collapse">
+  <div class="col-sm-5">
+<p style="text-align: justify;">Life Cycle Assessment is a systematic way of quantifying the environmental sustainability of a product and helps to identify aspects for improvement. It is a vital tool for sustainability studies and if implemented correctly can act as the basis to evaluate different scenarios and facilitate decisions.</p>
+<p><strong>Implementations</strong></p>
+<p style="text-align: justify;">The LCA resulted in an iterative tool that began as the repercussion of the dialogues with experts and ended as a decision making instrument. The LCA supported the idea of using the in-situ secretion system and showed us that the recycling scenario modelled for our process contributed to decrease the environmental impacts up to 70%. To read more about our LCA results please click <a href="https://2018.igem.org/Team:Edinburgh_OG/life_cycle_assessment"> <strong>here.</strong></a></p>
 </div>
 </div>
-  <div class="container"><button class="btn btn-primary" type="button" data-toggle="collapse" data-target="#tittle25">
-<p><strong>Week 9. Determination of PHA production </strong></p></button>
-        <div id="tittle25" class="collapse">
-<ul>
-<li>Nile red plate staining</li>
-<p>&nbsp;<img style="display: block; margin-left: auto; margin-right: auto;" src="https://static.igem.org/mediawiki/2018/4/4e/T--Edinburgh_OG--Notebook_-_bktb_6.png" width="425" height="214" /></p>
-<p style="text-align: center;"><strong>Figure 6</strong>. Nile red Plate staining of cells harbouring pSB1C3-phaCAB-bktB or pSB1C3-phaCB-bktB. Compared with control cells harbouring control plasmid, the production of PHA was confirmed.</p>
-<p>&nbsp;</p>
-<li>Extraction of produced PHA</li>
-<table style="margin-left: auto; margin-right: auto;" width="0">
-<tbody>
-<tr>
-<td style="width: 198px;">&nbsp;</td>
-<td style="width: 57px;">
-<p><strong>OD<sub>600</sub></strong></p>
-</td>
-<td style="width: 104px;">
-<p><strong>Volume (ml)</strong></p>
-</td>
-<td style="width: 113px;">
-<p><strong>Total PHA (g)</strong></p>
-</td>
-<td style="width: 104px;">
-<p><strong>PHA (mg/ml)</strong></p>
-</td>
-</tr>
-<tr>
-<td style="width: 198px;">
-<p><strong>pSB1C3</strong></p>
-</td>
-<td style="width: 57px;">
-<p>1.35</p>
-</td>
-<td style="width: 104px;">
-<p>100ml</p>
-</td>
-<td style="width: 113px;">
-<p>0</p>
-</td>
-<td style="width: 104px;">
-<p>0</p>
-</td>
-</tr>
-<tr>
-<td style="width: 198px;">
-<p><strong>pSB1C3-phaCAB</strong></p>
-</td>
-<td style="width: 57px;">
-<p>1.707</p>
-</td>
-<td style="width: 104px;">
-<p>100ml</p>
-</td>
-<td style="width: 113px;">
-<p>0.044</p>
-</td>
-<td style="width: 104px;">
-<p>0.44</p>
-</td>
-</tr>
-<tr>
-<td style="width: 198px;">
-<p><strong>pSB1C3-phaCB-bktB</strong></p>
-</td>
-<td style="width: 57px;">
-<p>2.15</p>
-</td>
-<td style="width: 104px;">
-<p>100ml</p>
-</td>
-<td style="width: 113px;">
-<p>0.023</p>
-</td>
-<td style="width: 104px;">
-<p>0.23</p>
-</td>
-</tr>
-<tr>
-<td style="width: 198px;">
-<p><strong>pSB1C3-phaCAB-bktB</strong></p>
-</td>
-<td style="width: 57px;">
-<p>1.875</p>
-</td>
-<td style="width: 104px;">
-<p>100ml</p>
-</td>
-<td style="width: 113px;">
-<p>0.021</p>
-</td>
-<td style="width: 104px;">
-<p>0.21</p>
-</td>
-</tr>
-</tbody>
-</table>
-<p>&nbsp;</p>
-<p>&nbsp;</p>
-<li>Measure the melting point of extracted PHA</li>
-<table style="height: 247px; margin-left: auto; margin-right: auto; width: 568px;">
-<tbody>
-<tr>
-<td style="width: 294.5px;">&nbsp;</td>
-<td style="width: 84.5px;"><strong>T<sub>m</sub> 1(&deg;C</strong><strong>)</strong></td>
-<td style="width: 85px;"><strong>T<sub>m</sub> 2 (&deg;C</strong><strong>)</strong></td>
-<td style="width: 84px;"><strong>T<sub>m</sub> 3 (&deg;C</strong><strong>)</strong></td>
-</tr>
-<tr>
-<td style="width: 294.5px;"><strong>Pure PHB from Sigma</strong></td>
-<td style="width: 84.5px;">170-179</td>
-<td style="width: 85px;">168-176</td>
-<td style="width: 84px;">168-174</td>
-</tr>
-<tr>
-<td style="width: 294.5px;"><strong>PHBV with 12% 3HV from Sigma</strong></td>
-<td style="width: 84.5px;">159-161</td>
-<td style="width: 85px;">160-160</td>
-<td style="width: 84px;">161-164</td>
-</tr>
-<tr>
-<td style="width: 294.5px;"><strong>PHA from pSB1C3-PhaCAB</strong></td>
-<td style="width: 84.5px;">160-168</td>
-<td style="width: 85px;">160-162</td>
-<td style="width: 84px;">161-164</td>
-</tr>
-<tr>
-<td style="width: 294.5px;"><strong>PHA from pSB1C3-PhaCB-bktB</strong></td>
-<td style="width: 84.5px;">150-155</td>
-<td style="width: 85px;">149-151</td>
-<td style="width: 84px;">149-152</td>
-</tr>
-<tr>
-<td style="width: 294.5px;"><strong>PHA from pSB1C3-PhaCAB-bktB</strong></td>
-<td style="width: 84.5px;">155-159</td>
-<td style="width: 85px;">156-161</td>
-<td style="width: 84px;">157-159</td>
-</tr>
-</tbody>
-</table>
-<p>&nbsp;</p>
-<p>Melting temperature of extracted PHA was measured and compared with PHB and PHBV products which were brought from Sigma. Tm of standard pure PHB product was between 170 &deg;C and 180 &deg;C and Tm of PHBV (12 % 3HV) was between 160 &deg;C and 164 &deg;C.The melting temperature of extracted PHA differed from different constructs. The PHA extracted from cell harbouring pSB1C3-phaCAB plasmids showed higher melting temperature with approximately 165 &deg;C. While the PHA extracted from the cells harbouring pSB1C3-phaCB-bktB or pSB1C3-phaCAB-bktB plasmids presented lower melting temperature at range of 150 &deg;C -155 &deg;C and 158 &deg;C -162 &deg;C respectively.</p>
-<p>&nbsp;</p>
-<p>&nbsp;</p>
 </div>
 </div>
-<h2><strong>Investigation of the effect of phasin autoregulation system on PHB production (Qihui)</strong></h2>
-  <div class="container"><button class="btn btn-primary" type="button" data-toggle="collapse" data-target="#tittle31">
-<p><strong>Week 1 Experimental preparation </strong></p></button>
-        <div id="tittle31" class="collapse">
-<ul>
-<li>Recover BioBricks from iGEM 2018 distribution kit</li>
-<li>Design the construct strategies. Two standard BioBricks (<strong>Table 1</strong>) were decided to be used for the experiments.</li>
-<p><strong>Table 1 </strong>Basic information of BioBricks</p>
-<table style="margin-left: auto; margin-right: auto;" width="0">
-<tbody>
-<tr>
-<td width="116">
-<p style="text-align: center;"><strong>Biobrick</strong></p>
-</td>
-<td style="text-align: center;" width="55">
-<p><strong>Well</strong></p>
-</td>
-<td style="text-align: center;" width="38">
-<p><strong>Plate</strong></p>
-</td>
-<td style="text-align: center;" width="108">
-<p><strong>Insert Length</strong></p>
-</td>
-<td style="text-align: center;" width="79">
-<p><strong>Backbone</strong></p>
-</td>
-<td width="162">
-<p style="text-align: center;"><strong>Antibiotics Resistance</strong></p>
-</td>
-</tr>
-<tr>
-<td width="116">
-<p><strong>BBa_K390501</strong></p>
-</td>
-<td style="text-align: center;" width="55">
-<p>19H</p>
-</td>
-<td style="text-align: center;" width="38">
-<p>6</p>
-</td>
-<td style="text-align: center;" width="108">
-<p>1136 bp</p>
-</td>
-<td style="text-align: center;" width="79">
-<p>pSB1C3</p>
-</td>
-<td style="text-align: center;" width="162">
-<p>chloramphenicol</p>
-</td>
-</tr>
-<tr>
-<td width="116">
-<p><strong>BBa_K1149051</strong></p>
-</td>
-<td style="text-align: center;" width="55">
-<p>12M</p>
-</td>
-<td style="text-align: center;" width="38">
-<p>4</p>
-</td>
-<td style="text-align: center;" width="108">
-<p>4271 bp</p>
-</td>
-<td style="text-align: center;" width="79">
-<p>pSB1C3</p>
-</td>
-<td style="text-align: center;" width="162">
-<p>chloramphenicol</p>
-</td>
-</tr>
-</tbody>
-</table>
-<p>&nbsp;</p>
-<li>Making competent cells (<em> coli</em> BL21 and DH5&alpha;)</li>
-<li>Double digestion for confirming the Biobricks</li>
-<li>Transformation test</li>
-</ul>
 </div>
 </div>
@@ Line 864: / Line 260: @@
-  <div class="container"><button class="btn btn-primary" type="button" data-toggle="collapse" data-target="#tittle32">
-<p><strong>Week 2 Designing the fragments and primers</strong></p></button>
-        <div id="tittle32" class="collapse">
-<ul>
-<li>Design and order fragments from IDT</li>
-<p><em>phaR</em> promoter-<em>phaR-phaP</em> promoter (<em>proR-phaR-proP</em>) and primers for amplifying DNA fragment of <em>phaP</em> and <em>HlyA</em></p>
-<li>Confirm the cloning strategy</li>
-<p style="text-align: center;"><strong><img style="display: block; margin-left: auto; margin-right: auto;" src="https://static.igem.org/mediawiki/2018/1/13/T--Edinburgh_OG--Notebook_-_Q1.png" />Figure1. Strategy of R-P-H assembly</strong></p>
-<li>Due to the ordered sequence with high content of GC, the first IDT order failed to complete. The sequence of IDT-synthesized DNA was optimized and reordered.</li>
-</div>
-</div>
+<!--5.  Cambridge -->
-  <div class="container"><button class="btn btn-primary" type="button" data-toggle="collapse" data-target="#tittle33">
-<p><strong>Week 3 &ndash;Week 5 Constructs establishment </strong></p></button>
-        <div id="tittle33" class="collapse">
-<li>Double digestion of backbone vector with restriction enzymes of <em>Spe</em>I and <em>Pst</em></li>
-<li>Gel electrophoresis and Gel extraction</li>
-<li>Amplify <em>phaP</em> and <em>phaP-HlyA </em>followed by gel electrophoresis</li>
+    <div class="container"><button class="btn btn-warning" type="button" data-toggle="collapse" data-target="#tittle6"><h3><strong> &#32; &nbsp;&nbsp; &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; &nbsp;Cambridge Consultants&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </strong></h3></button>
-<p>&nbsp;&nbsp;&nbsp;<img style="display: block; margin-left: auto; margin-right: auto;" src="https://static.igem.org/mediawiki/2018/8/82/T--Edinburgh_OG--Notebook_-_Q2.png" width="397" height="180" /></p>
+        <div id="tittle6" class="collapse">
-<p style="text-align: center;"><strong>Figure 2 </strong>Images of 1% gel electrophoresis -&nbsp;<strong>A.</strong>1.0% agarose gel electrophoresis for PCR product of <em>phaP</em> and <em>phaP-HlyA</em>. Lane: 1. 1 kb ladder (molecular weight marker, NEB). 2. 25&mu;L PCR reaction for <em>phaP</em> (569 bp). 3. 25&mu;L PCR reaction for <em>phaP-HlyA</em> (759 bp). <strong>B</strong>. 1.0% agarose gel electrophoresis for PCR product of <em>proR-phaR-proP</em>. Lane: 1. 100 bp ladder (NEB). 2. 25&mu;L reaction of PCR amplification for <em>proR-phaR-proP</em> (853 bp).</p>
-<li>Ligate digested backbone pSB3T5 with <em>phaP </em>and digested backbone with <em>phaP-HlyA, </em>followed by transformation to <em> coli</em> DH5&alpha; competent cells.</li>
-<li>Order the new primers for sequencing the end of <em>phaP</em> DNA fragment to test whether the stop codon exists.</li>
+   <div class="col-sm-5">
-<li>Send plasmid pSB3T5-phaP-HlyA to Dundee Sequencing Services for Sanger sequencing.</li>
+<blockquote>
-<li>Ligate DNA fragments <em>proR-phaR-proP </em>with pSB3T5-phaP and pSB3T5-phaP-HlyA respectively. Double digest constructs for confirmation.</li>
-<p><strong><img style="display: block; margin-left: auto; margin-right: auto;" src="https://static.igem.org/mediawiki/2018/7/77/T--Edinburgh_OG--Notebook_-_Q3.png" width="256" height="194" /></strong></p>
+<p><strong>Dr. James Hallinan, Dr. Steve Thomas</strong> &amp; <strong>Dr. James Brown,</strong>&ndash; Experts in Synthetic Biology and Material Sciences, Cambridge Consultants </p>
-<p style="text-align: center;"><strong>Figure 3 </strong>Image of diagnostic digest of pSB3T5-phaP and pSB3T5-phaP-hlyA with <em>Nsi</em>I and <em>Pst</em>I. Lanes - 1. 1 kb ladder (molecular weight marker, NEB). 2 and 3. Potential pSB3T5-phaP digested with NsiI and PstI. 4 and 5. Potential pSB3T5-phaP-HlyA digested with <em>Nsi</em>I and <em>Pst</em>I.</p>
+<p class="cd-timeline-content">The interviewed experts endorsed our project and explained that PHAs represent an innovating example of the delivery of sustainable products through cutting edge technology. They suggested the group to focus on the properties, design and potential applications of PHAs instead of tackling the production price. Furthermore, they emphasised the manufacturing challenges of producing PHAs, which are the major hurdle preventing their wide scale adoption: the scalability.</p>
-<li>Design primers for deleting the stop codon in between the <em>phaP</em> and <em>HlyA</em>.</li>
-</ul>
+     </blockquote>
+<div class="container"><button class="btn btn-info" type="button" data-toggle="collapse" data-target="#Cambridge">Read more</button>
+<div id="Cambridge" class="collapse">
+  <div class="col-sm-5">
+<p><strong>Assessing Feedstock and Downstream Processing</strong></p>
+<p>It was also discussed the topic relating to the present obstacles in bioplastic production, according to them feedstock resources and downstream processing are the key problems that must be addressed immediately. Feedstock is an environmentally sustainable way to sequester carbon source. Additionally, the interviews added the attractiveness of using underutilised products from industries as raw materials, because they are more budget friendly since they are usually under-priced, however,&nbsp;it needs to be built on an effective logistics strategy in terms of the usage as well as the consistency of the components in the by-product itself. The downstream processing is one of the biggest bottlenecks in the process, for this reason having a method for delivering the bioplastic without the necessity of disrupting the cells would increase the efficiency of the overall production and potentially can lead to the implementation of continuous-batch fermentation which could run for more hours and the carbon source and the product can be simultaneously added and removed, respectively.</p>
+<strong>Implementation and Impacts in our project </strong>
+<p>First, the use of whisky by-products as raw material is supported by the consistency of their elements. Second, the logistics of using them can be more achievable as they are the product of a local industry. The in-situ secretion system constitutes a potential method for reducing the use of hazardous chemicals in the process hence reduces the environmental impact of the PHBV production. Moreover, the potential use of continuous-batch fermentation can contribute to the overall efficiency of the manufacture. The cost of the bioplastic production is <em>not</em> the most important factor, due to the necessity of shift from oil to bio-based raw material. We took their advice of looking more into the properties and we decided to only focus on PHBV production because PHBV is a PHA copolymer with a larger thermal processing window, thus less brittle than PHB.</p>
+</div>
+</div>
 </div>
 </div>
-  <div class="container"><button class="btn btn-primary" type="button" data-toggle="collapse" data-target="#tittle34">
-<p><strong>Week 6 Repeat the experiments and finish constructs establishment. </strong></p></button>
-        <div id="tittle34" class="collapse">
-<ul>
-<li>Mini-prep for plasmids R-P and R-P-HlyA followed by double digestion. (failed). Results were showed in figure 4a.</li>
-<li>Amplify pSB3T5-phaP-HlyA (del) and <em>R-P-HlyA</em> (del) for deleting the stop codon. Running 0.8% agarose gel (figure 4b)</li>
-<p><img style="display: block; margin-left: auto; margin-right: auto;" src="https://static.igem.org/mediawiki/2018/5/53/T--Edinburgh_OG--Notebook_-_Q4.png" width="441" height="197" /></p>
-<p style="text-align: center;"><strong>Figure 4&nbsp;</strong>Images of gel electrophoresis.<strong> a.</strong> Diognostic digest of R-P and <em>R-P-HlyA</em> with <em>EcoR</em>I and <em>Pst</em>I. Lanes: 1. 1 kb ladder (molecular weight marker, NEB). 2. Potential R-P digested with <em>EcoR</em>I and <em>Pst</em>I. 3. Potential R-P-H digested with <em>EcoR</em>I and <em>Pst</em>I. <strong>b</strong>. 1.0% Agarose gel electrophoresis for PCR product of plasmid pSB3T5-phaP-HlyA (del) and <em>R-P-HlyA</em> (del).Lane: 1. 1kb ladder (NEB). 2. 25&mu;L reaction of PCR amplification pSB3T5-phaP-HlyA. 3. 25&mu;L reaction of PCR amplification <em>R-P-HlyA</em>.</p>
-<li>Co-transform the plasmid of phaCAB operon and the construct R-P into <em> coli</em> BL21 competent cells. The recovered cells were spread on the LB agar plates containing 10 &mu;g/ml tetracycline and 25&mu;g/ml chloramphenicol. (failed)</li>
-<li>Double digest pSB3T5 backbone to remove the RFP in between the <em>EcoR</em>I and <em>Xba</em></li>
-<li>Transformation:</li>
-<p>Repeat the co-transformation of <em>phaCAB</em> operon and R-P with lower concentration of antibiotics (8 &mu;g/ml tetracycline and 20 &mu;g/ml chloramphenicol).&nbsp;</p>
-<p>Transform plasmid R-P into <em>E. coli</em> BL21 competent cells.</p>
-<p>Transform the ligation of pSB3T5 without RFP (RFP (-) into <em>E. coli</em> DH5&alpha; competent cells.</p>
-<li>Colony PCR and double digestion</li>
-<p><img style="display: block; margin-left: auto; margin-right: auto;" src="https://static.igem.org/mediawiki/2018/e/e8/T--Edinburgh_OG--Notebook_-_Q5.png" width="578" height="142" /></p>
-<p><strong>Figure 5 </strong>Image of Agarose gel electrophoresis.<strong> a. </strong>1.0% Agarose gel electrophoresis of colony PCR product for screening the positive construct of P. &nbsp;Lane: 1. 1kb ladder (Promega). 2-14. 25&mu;L reaction of colony PCR amplification for <em>proP-phaP</em> (bp) with different colony extraction. &nbsp;The positive construct was expected to show a band with size of 1067 bp (<em>proP-phaP</em> + extra sequence in between the VF and VR primers.). <strong>b.</strong> 1.0% Agarose gel electrophoresis of colony PCR product for screening the positive construct of R.</p>
-<li>Mini-prep for plasmid R-P-HlyA (del) and P and transformation.</li>
-</ul>
 </div>
 </div>
@@ Line 951: / Line 310: @@
-  <div class="container"><button class="btn btn-primary" type="button" data-toggle="collapse" data-target="#tittle35"><p><strong>Week 8 Transformation and colony PCR</strong></p></button>
-        <div id="tittle35" class="collapse">
-<ul>
-<li>Measure the optical density of preculture of the recombinant <em> coli</em> BL21 strains that harboured R-P, <em>phaCAB</em> operon+R-P, <em>phaCAB</em> operon+ RFP (-), pSB1C3 (without RFP) +RFP (-) and RFP (-)</li>
-<li>Transformation:</li>
-<p>Co-transform the <em>phaCAB</em> operon and <em>R-P-HlyA </em>(del) into <em>E. coli</em> BL21 competent cells</p>
-<p>Co-transform the <em>phaCAB</em> operon and P into <em>E. coli</em> BL21 competent cells.</p>
-<p>Transform the <em>R-P-HlyA</em> (del) into <em>E. coli</em> BL21 competent cells.</p>
-<p>Transform the P into <em>E. coli</em> BL21 competent cells.</p>
-<li>Colony PCR to fast screen the positive constructs of R.</li>
-</ul>
-<p><img style="display: block; margin-left: auto; margin-right: auto;" src="https://static.igem.org/mediawiki/2018/1/17/T--Edinburgh_OG--Notebook_-_Q6.png" width="353" height="181" /></p>
-<p style="text-align: center;"><strong>Figure 6&nbsp;</strong>Image of 1.0% Agarose gel electrophoresis of colony PCR product for screening the positive construct of R -&nbsp;Lane: 1. 1 kb ladder (Promega). 2-14. 25&mu;L reaction of colony PCR amplification for <em>proR-phaR</em> with different colony extraction.</p>
-<p>The positive construct was expected to show a band with size of 935 bp (<em>proR-phaR</em> + extra sequence in between the VF and VR primers.</p>
-</div>
-</div>
+<!--6.  Meet-up -->
+    <div class="container"><button class="btn btn-warning" type="button" data-toggle="collapse" data-target="#tittle7"><h3><strong>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;iGEM Northern UK Meet-up&nbsp;@&nbsp;U&nbsp;St&nbsp;Andrews&nbsp;&nbsp;
+&nbsp;&nbsp;&nbsp;</strong></h3></button>
+        <div id="tittle7" class="collapse">
+   <div class="col-sm-5">
+<blockquote>
+<p style="text-align: justify;">We were invited to the&nbsp; iGEM&nbsp;Northern Meet-up&nbsp;organised by the St Andrews&nbsp;iGEM&nbsp;team. This event was intended for the cooperation between teams as a space to share ideas, experiences and feedback for the projects. During the event we got feedback from professors and previews generations of&nbsp;iGEMers.</p>
+     </blockquote>
+<div class="container"><button class="btn btn-info" type="button" data-toggle="collapse" data-target="#Meet-up">Read more</button>
+<div id="Meet-up" class="collapse">
-   <div class="container"><button class="btn btn-primary" type="button" data-toggle="collapse" data-target="#tittle36"><p><strong>Week 9&mdash;Week 10. Culture bacteria to produce PHB </strong></p></button>
-        <div id="tittle36" class="collapse">
+  <div class="col-sm-5">
+<p><strong>Implementation</strong></p>
+<p style="text-align: justify;">They gave us advice of how to construct our narrative and to shape our objectives.&nbsp;Regarding the design of the process, they suggested to&nbsp;look into&nbsp;the use of fed-batch or continuous fermentation in order to increase the yield of PHBV. Furthermore, they also suggested alternatives for our&nbsp;<em>in silico</em>&nbsp;model.</p>
+</div>
-<ul>
+</div>
-<li>Pre-culture all the recombinant <em> coli</em> BL21 strains: <em>phaCAB</em> <em>operon+R-P</em>, phaCAB <em>operon+R-P-HlyA</em> (del), <em>phaCAB operon+p</em>, <em>phaCAB operon+R</em>, <em>phaCAB operon+RFP(-), 1C3-RFP(-)+RFP(-).</em></li>
+</div>
-<li>The Nile red plate staining</li>
+</div>
-<p><img style="display: block; margin-left: auto; margin-right: auto;" src="https://static.igem.org/mediawiki/2018/9/9b/T--Edinburgh_OG--Notebook_-_Q7.png" width="390" height="272" /></p>
-<p style="text-align: center;"><strong>Figure 7&nbsp;</strong>Nile red plates to confirm PHB production (48 hours)</p>
-<p style="text-align: justify;">In each picture of a, b, c, d, e, there were 4 Nile red plates. The left plates in the first rows were strains harbouring <em>phaCAB</em> operon and constructs. The right plates in the first row were strains containing the construct only. The left plates in the second row were strains harbouring <em>phaCAB</em> operon only. The right plates in the second row were pSB1C3 and pSB3T5 backbones. <em>phaCAB</em> operon strain showed strong signal of fluorescence because of PHB produced and the backbone strain did not due to no PHB produced.</p>
-<li>Inoculate the preculture into 250 ml flasks with 50 ml of M9 medium containing 3.0% glucose, 25 &mu;g/ml chloramphenicol and 10 &mu;g/ml tetracycline.</li>
-<li>Measure the optical density to plot the growth curves.</li>
-<p>&nbsp;<img style="display: block; margin-left: auto; margin-right: auto;" src="https://static.igem.org/mediawiki/2018/6/65/T--Edinburgh_OG--Notebook_-_Q8.png" width="375" height="321" /></p>
-<p style="text-align: center;"><strong>Figure 7 </strong>The growth curves of E. coli strains that harboured different recombinant constructs</p>
-<p style="text-align: center;"><strong>Table 2 </strong>PHB production</p>
-<table style="margin-left: auto; margin-right: auto;" width="0">
-<tbody>
-<tr>
-<td width="208">&nbsp;</td>
-<td width="123">
-<p style="text-align: center;"><strong>Intracellular PHB</strong></p>
-</td>
-<td style="text-align: center;" width="102">
-<p><strong>Secreted PHB </strong></p>
-</td>
-<td style="text-align: center;" width="103">
-<p><strong>Mass of CaCl2</strong></p>
-</td>
-<td width="99">
-<p style="text-align: center;"><strong>Secreted PHB</strong></p>
-</td>
-</tr>
-<tr>
-<td width="208">
-<p><strong>phaCAB operon</strong><strong>+pSB3T5-R-P</strong></p>
-</td>
-<td style="text-align: center;" width="123">
-<p>0.071 g</p>
-</td>
-<td style="text-align: center;" width="102">
-<p>0.057 g</p>
-</td>
-<td style="text-align: center;" width="103">
-<p>0.05549 g</p>
-</td>
-<td style="text-align: center;" width="99">
-<p>0.0015 g</p>
-</td>
-</tr>
-<tr>
-<td width="208">
-<p><strong>phaCAB operon</strong><strong>+pSB3T5-R-P-H</strong></p>
-</td>
-<td style="text-align: center;" width="123">
-<p>0.043 g</p>
-</td>
-<td style="text-align: center;" width="102">
-<p>0.086 g</p>
-</td>
-<td style="text-align: center;" width="103">
-<p>0.05549 g</p>
-</td>
-<td style="text-align: center;" width="99">
-<p>0.0305 g</p>
-</td>
-</tr>
-<tr>
-<td width="208">
-<p><strong>phaCAB operon</strong><strong>+pSB3T5-R</strong></p>
-</td>
-<td style="text-align: center;" width="123">
-<p>0.065 g</p>
-</td>
-<td style="text-align: center;" width="102">
-<p>0.058 g</p>
-</td>
-<td style="text-align: center;" width="103">
-<p>0.05549 g</p>
-</td>
-<td style="text-align: center;" width="99">
-<p>0.0020 g</p>
-</td>
-</tr>
-<tr>
-<td width="208">
-<p><strong>phaCAB operon +pSB3T5-P</strong></p>
-</td>
-<td style="text-align: center;" width="123">
-<p>0.031 g</p>
-</td>
-<td style="text-align: center;" width="102">
-<p>0.030 g</p>
-</td>
-<td style="text-align: center;" width="103">
-<p>0.05549 g</p>
-</td>
-<td style="text-align: center;" width="99">
-<p>0.0000 g</p>
-</td>
-</tr>
-<tr>
-<td width="208">
-<p><strong>phaCAB operon +pSB3T5</strong></p>
-</td>
-<td style="text-align: center;" width="123">
-<p>0.010 g</p>
-</td>
-<td style="text-align: center;" width="102">
-<p>0.038 g</p>
-</td>
-<td style="text-align: center;" width="103">
-<p>0.05549 g</p>
-</td>
-<td style="text-align: center;" width="99">
-<p>0.0000 g</p>
-</td>
-</tr>
-<tr>
-<td width="208">
-<p><strong>pSB1C3+pSB3T5</strong></p>
-</td>
-<td style="text-align: center;" width="123">
-<p>0.000 g</p>
-</td>
-<td style="text-align: center;" width="102">
-<p>0.042 g</p>
-</td>
-<td style="text-align: center;" width="103">
-<p>0.05549 g</p>
-</td>
-<td style="text-align: center;" width="99">
-<p>0.0000 g</p>
-</td>
-</tr>
-</tbody>
-</table>
-<li>Extract produced PHB (The dry weight of extracted and secreted PHB were listed in the Table 2</li>
-<p style="text-align: center;"><strong><img src="https://static.igem.org/mediawiki/2018/f/f8/T--Edinburgh_OG--Notebook_-_Q9.png" width="543" height="293" /></strong></p>
-<p style="text-align: center;"><strong>Figure 8 </strong>Column graph for dry weight of secreted and intracellular PHB among different stains</p>
-<li>Measure the melting point of produced PHB.</li>
-</ul>
-<p style="text-align: center;"><strong>Table 3 </strong>The melting temperature of produced PHB</p>
-<table style="margin-left: auto; margin-right: auto;" width="0">
-<tbody>
-<tr>
-<td width="273">
-<p><strong>Strains</strong></p>
-</td>
-<td width="294">
-<p><strong>Melting Temperature</strong></p>
-</td>
-</tr>
-<tr>
-<td width="273">
-<p><strong>phaCAB operon+pSB3T5-R-P</strong></p>
-</td>
-<td style="text-align: center;" width="294">
-<p>156-164&deg;C</p>
-</td>
-</tr>
-<tr>
-<td width="273">
-<p><strong>phaCAB operon+pSB3T5-R-P-H</strong></p>
-</td>
-<td style="text-align: center;" width="294">
-<p>160-168&deg;C</p>
-</td>
-</tr>
-<tr>
-<td width="273">
-<p><strong>phaCAB operon +pSB3T5-R</strong></p>
-</td>
-<td style="text-align: center;" width="294">
-<p>156-164&deg;C</p>
-</td>
-</tr>
-<tr>
-<td width="273">
-<p><strong>phaCAB operon +pSB3T5-P</strong></p>
-</td>
-<td style="text-align: center;" width="294">
-<p>152-164&deg;C</p>
-</td>
-</tr>
-<tr>
-<td width="273">
-<p><strong>phaCAB operon+pSB3T5</strong></p>
-</td>
-<td style="text-align: center;" width="294">
-<p>162-170&deg;C</p>
-</td>
-</tr>
-<tr>
-<td width="273">
-<p><strong>pSB1C3+pSB3T5</strong></p>
-</td>
-<td style="text-align: center;" width="294">
-<p>\</p>
-</td>
-</tr>
-</tbody>
-</table>
-<p>&nbsp;</p>
-<p>&nbsp;</p>
 </div>
 </div>
@@ Line 1,200: / Line 360: @@
-<h2><strong>Construction of Plasmids Harbouring the Sleeping Beauty Mutase Operon and Methylmalonyl CoA Epimerase for Producing Propionate for PHBV Production (Craig)</strong></h2>
+<!--7.  Whisky association-->
@@ Line 1,209: / Line 369: @@
-  <div class="container"><button class="btn btn-primary" type="button" data-toggle="collapse" data-target="#tittle41"><p><strong>Week 1 &ndash; Week 2</strong></p></button>
-         <div id="tittle41" class="collapse">
+    <div class="container"><button class="btn btn-warning" type="button" data-toggle="collapse" data-target="#tittle8"><h3><strong>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Scotch Whisky Association&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</strong><span data-ccp-props="{&quot;201341983&quot;:0,&quot;335551550&quot;:6,&quot;335551620&quot;:6,&quot;335559739&quot;:160,&quot;335559740&quot;:259}">&nbsp;</span></h3></button>
+         <div id="tittle8" class="collapse">
+   <div class="col-sm-5">
+<blockquote>
+<p style="text-align: justify;"><strong>Morag Garden</strong>&nbsp;- Head of Sustainability &amp; Innovation, Scotch Whisky Association</p>
+<p style="text-align: justify;">Morag facilitated the group a lot of information about the local situation of the whisky market and their by-products. She highlighted the increasing global demand for Scotch and the now 128 distillers operating across Scotland's five different&nbsp;whisky&nbsp; regions. In addition, she gave us suggestions for alternative community members looking at valorising the distillery materials, as they would be a useful source of information.&nbsp; Additionally, she also mentioned their interest in the outcomes of the research.</p>
+     </blockquote>
+<div class="container"><button class="btn btn-info" type="button" data-toggle="collapse" data-target="#Scotch">Read more</button>
+<div id="Scotch" class="collapse">
+  <div class="col-sm-5">
+<p><strong>Some notes from the Whisky by-products study case</strong></p>
+<p style="text-align: justify;">The whisky sector remains high with a growth rate estimated at 2.1% over the period to 2018. This would result in an approximate increase in by-products in the region of:&nbsp;Draff&nbsp;&ndash; 52,800 tonnes and Pot ale &ndash; 88,800 tonnes. By-products from Scotch Whisky distillery (draff&nbsp;and pot ale for example), have long been reused by the agriculture sector as a valuable animal feed and fertiliser for agricultural land (Figure 1). However, suitable routes would be need to be found for these by-products, as existing are near their natural capacity. New developments have recently&nbsp;opened up&nbsp;new markets for the use of by-products as feedstock for renewable heat or its use to make protein feed for salmon farming and biofuel.&nbsp;</p>
+<p style="text-align: center;"><img src="https://static.igem.org/mediawiki/2018/e/e3/T--Edinburgh_OG--IHP_-_A.png" width="565" height="311" /></p>
+<p style="text-align: center;"><strong>Figure 1</strong>&nbsp;Availability of by-product feedstocks by local authorities in Scotland. Taken from <strong>Ricardo Energy &amp; Environment for Zero Waste Scotland (2017). Biorefining Potential for Scotland.</strong>&nbsp;</p>
+<p><img style="display: block; margin-left: auto; margin-right: auto;" src="https://static.igem.org/mediawiki/2018/7/7f/T--Edinburgh_OG--IHP_-_B.png" width="545" height="357" /></p>
+<p style="text-align: center;">Figure 2. The current uses for whisky by-products in Scotland. Taken from <strong>Zero Waste Scotland (2015). Sector Study on Beer, Whisky and Fish. Final Report</strong></p>
+<p style="text-align: justify;">A circular economy approach is to use resources more efficiently, keep resources in use for as long a time as possible and to minimise waste. Identifying opportunities to improve circular approaches, for example keeping by-product materials within Scotland for further processing. There are many potential products that could feasibly be made from by-products, adding further value to a business.</p>
+<p style="text-align: justify;"><strong>References</strong></p>
 <ul>
-<li>Discuss the project aims and decide individual project strategy (shown in Figure 1).</li>
+<li>Miller, R. The landscape for biopolymers in packaging. Miller- Klein Associates report Summary and Full Report available from The National Non-Food Crops Centre, Heslington, New York, UK, 2005, <a href="http://www.nnfcc.co.uk">nnfcc.co.uk</a>.</li>
-<p><strong><img style="display: block; margin-left: auto; margin-right: auto;" src="https://static.igem.org/mediawiki/2018/b/b7/T--Edinburgh_OG--PHBV_prodxn_-_5.png" width="527" height="305" /></strong></p>
+<li>Scottish Environment Protection Agency (2018). Scotch Whisky Sector Plan. Available at: <a href="https://consultation.sepa.org.uk/communications/sector-approach-to-regulation-consultations-on-sco/supporting_documents/SEPA_Whisky%20Sector%20Plan_Final.pdf">https://consultation.sepa.org.uk/communications/sector-approach-to-regulation-consultations-on-sco/supporting_documents/SEPA_Whisky%20Sector%20Plan_Final.pdf</a></li>
-<p style="text-align: center;"><strong>Figure 1 </strong>Proposed mechanism for propionate synthesis utilising the Sleeping beauty mutase operon (SBM) and Methylmalonyl-CoA epimerase (MCE) -&nbsp;Succinyl-CoA is converted into Methylmalonyl-CoA-R by the methylmalonyl- CoA mutase ScpA. Methylmalonyl-CoA-R is converted into Methylmalonyl- CoA-S by MCE or an uncharacterised, native pathway. Methylmalonyl-CoA-S is converted into propionyl-CoA by the methylmalonyl-CoA carboxylase ScpB. The CoA from Propionyl-CoA is transferred onto Succinate from the citric acid cycle by the Propionyl-CoA: Succinate CoA transferase ScpC, resulting in the production of propionate and Succinyl-CoA.</p>
+<li>Ricardo Energy &amp; Environment for Zero Waste Scotland (2017). Biorefining Potential for Scotland. Available at: <a href="https://www.zerowastescotland.org.uk/sites/default/files/Biorefining%20Potential%20for%20Scotland%20Final%20report.pdf">https://www.zerowastescotland.org.uk/sites/default/files/Biorefining%20Potential%20for%20Scotland%20Final%20report.pdf</a></li>
-<li>Determine whether <em>E. coli</em>&nbsp; DH5 proposes SBM in genome and if it proposes any change that impact the resulting protein structure and function.</li>
-<li>Design and order DNA fragments from IDT to obtain Sleeping Beauty Mutase (SBM) which consists of genes <em>ScpA</em>, <em>ScpB</em>, <em>ScpC</em>, and</li>
-<li>Design and order the primers for amplifying SBM operon.</li>
 </ul>
-<p><img style="display: block; margin-left: auto; margin-right: auto;" src="https://static.igem.org/mediawiki/2018/8/87/T--Edinburgh_OG--Notebook_-_C2.png" width="638" height="332" /></p>
-<p style="text-align: center;"><strong>Figure 2 </strong>Illustrated diagrams of designed primers -&nbsp;A) The combined primers of Sbm Forward 1 and 2. Sbm Forward 1 annealed to <em>ScpA</em> at its 3&rsquo; end, and introduces a Ribosome binding site (RBS; green), <em>AvrI</em>I restriction site (black), and half of Promoter BBa_J23110 (orange). Sbm Forward 2 annealed to the amplified sbm operon through containing a complete Promoter BBa_J23110 sequence, and introduced the remaining sequence, alongside <em>Xba</em>I (blue) and <em>EcoR</em>I (red) restriction sites. B) Illustrated diagram of Sbm Reverse Primer. Anneals to the end of<em> ScpC</em> (green), and introduces a <em>Spe</em>I restriction site (orange), and a <em>Nsi</em>I restriction site (red).</p>
 </div>
 </div>
+</div>
+</div>
-  <div class="container"><button class="btn btn-primary" type="button" data-toggle="collapse" data-target="#tittle42"><p><strong>Week 3 &ndash; Week 5 Amplify SBM and construct cloning vectors. </strong></p></button>
-        <div id="tittle42" class="collapse">
-<ul>
-<li>Amplify the SBM operon with ordered primers as shown in Figure 1 by tow-step PCR. And analysed by gel electrophoresis.</li>
-<p><strong><img style="display: block; margin-left: auto; margin-right: auto;" src="https://static.igem.org/mediawiki/2018/3/37/T--Edinburgh_OG--Notebook_-_C3.png" width="284" height="430" /></strong></p>
-<p style="text-align: center;"><strong>Figure 3&nbsp;</strong>Gel Electrophoresis of DNA from 2 PCR reactions - PCR-1 is PCR results from the amplification of the SBM operon from E. coli DH5-gDNA, with a band between 6kbp and 5kbp, and a band of below 500bp. PCR-2 used the products from PCR-1 as template DNA and used the Sbm forward 2 and Sbm Reverse primers &ndash; a single band of below 500bp can be observed.</p>
-<li>TA cloning with SBM to obtain more SBM operon without doing PCR many times.</li>
-<p>SBM operon is &lsquo;A-tailed&rsquo; prior to ligating into pCRTM2.1-TOPO, and transformed into One Shot&reg; cells and plated. Following incubating overnight, blue colonies and white colonies could be observed. White colonies were used to purify plasmid DNA, and were analysed by digestion with <em>EcoR</em>I.</p>
-<p><strong><img style="display: block; margin-left: auto; margin-right: auto;" src="https://static.igem.org/mediawiki/2018/9/9f/T--Edinburgh_OG--Notebook_-_C4.png" width="480" height="274" /></strong></p>
-<p style="text-align: center;"><strong>Figure 4. Analytical digest of TA colonies.</strong> Marker ladder used was NEB 1kb ladder, and all samples show a band between 3kbp and 4kbp, and a band below 500bp.</p>
-<p style="text-align: justify;">Bands of below 500bp could be observed, suggesting that some of the primer dimers may have been contaminating the extracted SBM operon sample, which were A-tailed, and effectively out- competing the much larger SBM operon fragments to be ligated into pCRTM2.1-TOPO.Again, another cloning method for the utilisation of the SBM operon had to be devised.</p>
-<li>Construct pSB3T5: MCE: Promoter- RFP and pSB3T5: Promoter-RFP followed by gel electrophoresis.</li>
-<p style="text-align: center;"><strong><img src="https://static.igem.org/mediawiki/2018/b/b5/T--Edinburgh_OG--Notebook_-_C5.png" width="575" height="328" /></strong></p>
-<p style="text-align: center;"><strong>Figure 5 </strong>Restriction Analysis of various constructs. Marker ladder was Promega 1kb ladder. pSB3T5: MCE: Promoter-RFP was digested with <em>Spe</em>I and/or <em>Pst</em>I. Single digests both produced bands of 4757bp, whereas double digest produced 2 bands of 887bp and 3870bp. pSB3T5 was digested with <em>EcoR</em>I and/or <em>Pst</em>I. Single digestion results in bands of 4280bp, whereas double digest produced bands of 3211bp and 1069bp. pSB3T5: Promoter-RFP was digested with SpeI and/or PstI. Single digests both produced bands of 4156bp, whereas double digest produced 2 bands of about 3269bp, and 887bp. BBa_J23106 was digested with <em>EcoR</em>I and/or <em>Pst</em>I. Single digestion results in bands of 2983bp, whereas double digest produced bands of 2038bp, and 945bp. PSB3T5: MCE was digested with <em>EcoR</em>I and/or <em>Pst</em>I. Single digests both produced bands of 3845bp, whereas double digest produced 2 bands of 3211bp and 634bp.</p>
-<li>Insert the operon into BioBrick vevtor such as pSB3T5, through digesting with <em>EcoR</em>I and <em>Nsi</em></li>
-</ul>
-<p><img style="display: block; margin-left: auto; margin-right: auto;" src="https://static.igem.org/mediawiki/2018/2/21/T--Edinburgh_OG--Notebook_-_C6.png" width="854" height="227" /></p>
-<p><strong>Figure 6</strong>&nbsp;<strong>a.</strong> Diagram illustrating the construction of pSB3T5: MCE. RFP (red) was cleaved from pSB3T5 by digestion with <em>EcoR</em>I and PstI. <strong>b.</strong> Diagram illustrating the construction of pSB3T5: Promoter-RFP. RFP (red) was excised from pSB3T5 by digesting <em>EcoR</em>I and <em>Pst</em>I.</p>
 </div>
 </div>
@@ Line 1,274: / Line 415: @@
-  <div class="container"><button class="btn btn-primary" type="button" data-toggle="collapse" data-target="#tittle43"><p><strong>Week 6- Week7 Construct pSB3T5: SBM and pSB3T5: MCE: SBM</strong></p></button>
-        <div id="tittle43" class="collapse">
+<!--8.  Visiting Local distilleries -->
-<ul>
-<li>Construct pSB3T5: SBM followed by Colony PCR. The strategy was showed in Figure 7 and gel electrophoresis of colony PCR was showed in Figure 8.</li>
-<p><img style="display: block; margin-left: auto; margin-right: auto;" src="https://static.igem.org/mediawiki/2018/e/e5/T--Edinburgh_OG--Notebook_-_C7.png" width="513" height="265" /></p>
+    <div class="container"><button class="btn btn-warning" type="button" data-toggle="collapse" data-target="#tittle9"><h3><strong>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Visit to Local Whisky Distillery in Edinburgh&nbsp;&nbsp;&nbsp;&nbsp;</strong><span data-ccp-props="{&quot;201341983&quot;:0,&quot;335551550&quot;:6,&quot;335551620&quot;:6,&quot;335559739&quot;:160,&quot;335559740&quot;:259}">&nbsp;</span></h3></button>
-<p style="text-align: center;"><strong>Figure 7 </strong>Diagram illustrating the construction of pSB3T5 - SBM. pSB3T5: Promoter- RFP was digested with <em>Spe</em>I and <em>Pst</em>I, removing RFP (red). The SBM operon (orange) was digested with <em>AvrI</em>I and <em>Nsi</em>I. The overhangs generated are illustrated. The SBM operon was ligated into pSB3T5: Promoter downstream of the promoter (purple).</p>
+        <div id="tittle9" class="collapse">
-<p style="text-align: center;"><img src="https://static.igem.org/mediawiki/2018/a/ac/T--Edinburgh_OG--Notebook_-_C8.png" width="710" height="296" />&nbsp;&nbsp;</p>
-<p style="text-align: center;"><strong>Figure 8 a. </strong>Image of DNA gel electrophoresis from colony PCR of transformants possessing pSB3T5: SBM using SBM Forward 1 and SBM Reverse Primers. Control lane is the same PCR with no template DNA. Control shows a large empty lane except for a single band below 250bp. All samples show a large smear that is intense from the well down to 5551bp, corresponding to the SBM operon, where it becomes less intense. <strong>b.</strong> Analytical digest of pSB3T5: SBM using <em>Xba</em>I and <em>Spe</em>I, followed by gel electrophoresis. Uncut sample showed 2 bands, one above 10kbp, and one between 8kbp &ndash; 6kbp. <em>Xba</em>I and <em>Spe</em>I single digests show a single band of 8790bp. <em>Xba</em>I and <em>Spe</em>I double digest shows two bands: a pSB3T5 backbone at 3250bp, and the SBM operon at 5540bp.</p>
-<li>Two different strategies to construct pSB3T5: MCE: SBM</li>
+   <div class="col-sm-5">
-</ul>
+<blockquote>
-<p><img style="display: block; margin-left: auto; margin-right: auto;" src="https://static.igem.org/mediawiki/2018/d/d0/T--Edinburgh_OG--Notebook_-_C9.png" /></p>
-<p style="text-align: center;"><strong>Figure 9</strong>&nbsp;<strong>a.</strong> Diagram illustrating the construction of pSB3T5: MCE: SBM. pSB3T5: MCE: Promoter-RFP was digested with <em>Spe</em>I and <em>Pst</em>I, removing RFP (red). The SBM operon (orange) was digested with <em>AvrI</em>I and <em>Nsi</em>I. The overhangs generated are illustrated. The SBM operon was ligated into pSB3T5: Promoter downstream of the promoter (purple). MCE is indicated by black. <strong>b.</strong> Diagram illustrating the construction of pSB3T5: MCE: SBM. pSB3T5: SBM was digested with <em>EcoR</em>I and <em>Xba</em>I. pSB3T5: MCE was digested with <em>EcoR</em>I and <em>Spe</em>I to excise MCE (brown). The resulting overhangs are displayed. MCE was ligated into the plasmid upstream of the promoter (purple) and the SBM operon (orange), completing the plasmid.</p>
+<p style="text-align: justify;">Through this visit, we learnt about the whisky production and collected whisky production by-products samples (i.e. pot ale and draff) to be used as carbon source in our experiments.&nbsp;</p>
+     </blockquote>
+<div class="container"><button class="btn btn-info" type="button" data-toggle="collapse" data-target="#Whisky">Read more</button>
+<div id="Whisky" class="collapse">
+  <div class="col-sm-5">
+<p>We were introduced to the whisky production through the tour throughout the distillery. We learnt that the whisky industry managed their resources locally and sustainably and ensured that minimum waste is generated in every batch of the production. The information obtained was very helpful in conducting the Life Cycle Assessment.&nbsp;&nbsp;</p>
+<div class="collapse">&nbsp;</div>
+<div class="collapse" style="text-align: center;"><img src="https://static.igem.org/mediawiki/2018/a/a3/T--Edinburgh_OG--IHP_-_Cx.jpg" width="650" height="223" />&nbsp;<img src="https://static.igem.org/mediawiki/2018/f/f1/T--Edinburgh_OG--IHP_-_C.jpg" width="650" height="223" />&nbsp;<img src="https://static.igem.org/mediawiki/2018/b/b0/T--Edinburgh_OG--IHP_-_D.jpg" width="335" height="223" /></div>
+<div class="collapse" style="text-align: center;"><strong>Figure 1</strong> (from top to bottom) Tour guide explained about the kiln. Samples of malt before separated into husk, grist, and flour. Our team collected samples.&nbsp;</div>
+</div>
+</div>
+</div>
+</div>
 </div>
 </div>
@@ Line 1,303: / Line 458: @@
-  <div class="container"><button class="btn btn-primary" type="button" data-toggle="collapse" data-target="#tittle44"><p><strong>Week 8 Propionate Assay</strong></p></button>
-        <div id="tittle44" class="collapse">
-<ul>
-<li>Pellets of pSB3T5, pSB3T5: MCE, pSB3T5: SBM, and pSB3T5: MCE: SBM grown in LB broth were resuspended in M9 minimal media with 3% glucose and tetracycline in order to assay for the production of propionate.</li>
-<li>Detect the produced propionate by detecting the change in absorbance at 410nm using spectrophotometer.</li>
-<p>Fe<sup>3+</sup> ions have been shown to react with short chain fatty acids, such as propionate, where the ion complexes with the organic acid and is reduced. This changes the colour of the iron ion, and this change can be detected in a spectrophotometer by a change in absorbance at 410nm. (failed)</p>
-</ul>
-<p style="text-align: center;"><strong>Table 1 </strong>Absorbance at 410nm of propionate standard solutions</p>
-<table style="margin-left: auto; margin-right: auto;" width="586">
-<tbody>
-<tr>
-<td width="227">
-<p style="text-align: center;"><strong>Sample </strong></p>
-</td>
-<td width="359">
-<p style="text-align: center;"><strong>Absorbance at 410nm (arbitrary units) </strong></p>
-</td>
-</tr>
-<tr>
-<td width="227">
-<p><strong>0 g/l propionate </strong></p>
-</td>
-<td width="359">
-<p>0</p>
-</td>
-</tr>
-<tr>
-<td width="227">
-<p><strong>0.5 g/l propionate </strong></p>
-</td>
-<td width="359">
-<p>0.017</p>
-</td>
-</tr>
-<tr>
-<td width="227">
-<p><strong>1.0 g/l propionate </strong></p>
-</td>
-<td width="359">
-<p>0.003</p>
-</td>
-</tr>
-<tr>
-<td width="227">
-<p><strong>1.5 g/l propionate </strong></p>
-</td>
-<td width="359">
-<p>-0.008</p>
-</td>
-</tr>
-<tr>
-<td width="227">
-<p><strong>2.0 g/l propionate </strong></p>
-</td>
-<td width="359">
-<p>0.021</p>
-</td>
-</tr>
-<tr>
-<td width="227">
-<p><strong>2.5 g/l propionate </strong></p>
-</td>
-<td width="359">
-<p>0.002</p>
-</td>
-</tr>
-<tr>
-<td width="227">
-<p><strong>3.0 g/l propionate </strong></p>
-</td>
-<td width="359">
-<p>0.024</p>
-</td>
-</tr>
-<tr>
-<td width="227">
-<p><strong>3.5 g/l propionate </strong></p>
-</td>
-<td width="359">
-<p>0.026</p>
-</td>
-</tr>
-<tr>
-<td width="227">
-<p><strong>4.0 g/l propionate </strong></p>
-</td>
-<td width="359">
-<p>-0.003</p>
-</td>
-</tr>
-<tr>
-<td width="227">
-<p><strong>psb3T5 </strong></p>
-</td>
-<td width="359">
-<p>-0.5</p>
-</td>
-</tr>
-<tr>
-<td width="227">
-<p><strong>psb3T5: MCE</strong></p>
-</td>
-<td width="359">
-<p>-0.236</p>
-</td>
-</tr>
-<tr>
-<td width="227">
-<p><strong>psb3T5: SBM</strong></p>
-</td>
-<td width="359">
-<p>-0.232</p>
-</td>
-</tr>
-<tr>
-<td width="227">
-<p><strong>psb3T5: MCE: SBM</strong></p>
-</td>
-<td width="359">
-<p>-0.288</p>
-</td>
-</tr>
-</tbody>
-</table>
-<p>&nbsp;</p>
-<p>&nbsp;</p>
-</div>
+<!--9.  Bluepha -->
-</div>
@@ Line 1,444: / Line 472: @@
-<h2><strong>Investigation of the effects of <em>sucAB</em> and <em>sucCD</em> on the adaption to propionic Acid and PHBV production (Siqi)</strong></h2>
+    <div class="container"><button class="btn btn-warning" type="button" data-toggle="collapse" data-target="#tittle2"><h3><strong> &#32; &nbsp;  &nbsp; &nbsp;&nbsp;  &nbsp; &nbsp; &nbsp; &nbsp;  &nbsp; &nbsp; &nbsp; &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;BLUEPHA Technologies &nbsp; &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; &nbsp; &nbsp; &nbsp; &nbsp;  &nbsp; &nbsp; &nbsp;   &nbsp; &nbsp; </strong></h3></button>
+         <div id="tittle2" class="collapse">
-  <div class="container"><button class="btn btn-primary" type="button" data-toggle="collapse" data-target="#tittle51"><p><strong>Week 1- Week 2 Amplify essential DNA fragments by ordered primers</strong></p></button>
+   <div class="col-sm-5">
-         <div id="tittle51" class="collapse">
+<blockquote>
+<p><strong>Dr. Guo-Qiang Chen</strong> - Director of Center for Synthetic and Systems Biology, Tsinghua University and Chief Scientist at BLUEPHA</p>
+<p><strong>Jin Yin - </strong>Technique Manager at BLUEPHA</p>
-<ul>
+<p style="text-align: justify;">They endorsed our project explaining that our work is conducive to the long-term development of the world. As global environmental awareness increases, the market for biodegradable plastics will certainly develop. This includes the PHA market and research is needed to achieve this shift from plastics to bioplastics, especially in Europe where  composting action is encouraged by the government. PHAs certainly has this unique advantage over other bioplastics.&nbsp;</p>
-<li>Design and order primers from IDT</li>
+<p style="text-align: justify;">Over the interviews with Jin Yin and Dr. Chen, it was discussed the ways of solving the plastic pollution. In their point of view, one of them is promoting the use of bio-based plastics for containers and bags. However, they emphasised the necessity of research institutes and industry to work in the optimisation of the manufacturing processes and strain properties. For them, the bottlenecks in the PHA production are the strain properties, as the ability of using different raw materials, and the downstream process which is limiting the efficiency and purity of the product. As SynBio practitioners, they believe that the answer is in the use of this discipline to help solving these problems.</p>
-<li>Amplify the <em>sucAB</em> and <em>sucCD</em> from <em> coli</em> DH5genome (Figure 1),&nbsp;<em>sucAB</em> and <em>sucCD</em> are expected to be 4 kb and 2kb respectively.</li>
-</ul>
+     </blockquote>
-<p><strong><img style="display: block; margin-left: auto; margin-right: auto;" src="https://static.igem.org/mediawiki/2018/2/24/T--Edinburgh_OG--Notebook_-_S1.png" width="432" height="227" /></strong></p>
-<p style="text-align: center;"><strong>Figure 1&nbsp;</strong>Gel electrophoresis image of <em>sucAB</em> and <em>sucCD</em> genes from PCR amplification on 1% agarose gel, contrast with 1KB ladder from NEB, generating fragments for the following experiments.</p>
+<div class="container"><button class="btn btn-info" type="button" data-toggle="collapse" data-target="#BluePHA">Read more</button>
+<div id="BluePHA" class="collapse">
+  <div class="col-sm-5">
+<p style="text-align: justify;">Bluepha&nbsp;Co., Ltd. is a synthetic biology and biomaterial company established in Beijing, China. The company was the first SynBio Startup Founded by iGEMers, and they were the winner <em>of the Future Planet Award for Sustainable Growth</em>. Moreover, they are pioneering a revolutionary cost-effective method for producing bioplastics.</p>
+<p>Jin Yin and Dr. Chen:</p>
+<p><em>&ldquo;</em>Over 20 companies from different countries are focusing on the optimisation of PHA production, all of us are trying hard on solving many practical problems and we truly believe that the answer of some of these problems is in the use of Synthetic Biology. However, it is also important to focus on the discovery of medical applications of PHAs. The biocompatibility of PHAs enclose the potential for using it as artificial cartilage and nerve conducts. For example, US has permit the usage of P(4HB) in surgery. They believe that PHAs is a new thing deserving further studies with surprising properties that can be a potential answer for diseases such like osteoporosis. Despite of the current drawbacks, we believe in a more optimistic future for PHAs<em>&ldquo;</em><p>
+   <p>&nbsp;</p>
+<p><strong>Implementations</strong></p>
+<p style="text-align: justify;">We confirmed that our in-situ secretion system is a good approach to tackle the downstream processing problem. As further directions, they recommended us to look for improvements to the strain for being able to use whisky by-products more efficiently. However, it is important to note, that solving technical issues of PHA production is as important as it is related to the biomedical research. As a result, a new focus for our future research is to look into new biomedical applications of PHAs and specifically PHBV.</p>
 </div>
+  </div>
 </div>
+        </div>
+      </div>
+         </div>
+<!--10. SINOPEC -->
-  <div class="container"><button class="btn btn-primary" type="button" data-toggle="collapse" data-target="#tittle52"><p><strong>Week 3 &ndash; Week 5 Constructs establishment</strong></p></button>
-         <div id="tittle52" class="collapse">
+    <div class="container"><button class="btn btn-warning" type="button" data-toggle="collapse" data-target="#tittle10"><h3><strong>&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;SINOPEC&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; </em></strong></h3></button>
+         <div id="tittle10" class="collapse">
-<ul>
-<li>Construct plasmids: pSB3T5-sucAB, pSB3T5-sucCD, plasmid pSB3T5-X</li>
-<li>Transform recombinant plasmids to <em> coli</em> DH5</li>
+   <div class="col-sm-5">
-<li>Isolate the plasmids and double digest them with corresponding restriction enzymes for confirmation</li>
+<blockquote>
-</ul>
-<p><strong><img style="display: block; margin-left: auto; margin-right: auto;" src="https://static.igem.org/mediawiki/2018/2/28/T--Edinburgh_OG--Notebook_-_S2.png" width="576" height="232" /></strong></p>
+<p><strong>Paul Tan</strong> &ndash; Officer SINOPEC Guangzhou, China</p>
-<p style="text-align: center;"><strong>Figure 2. A</strong> Gel electrophoresis image of pSB3T5-sucAB digested with <em>EcoR</em>I and <em>Hind</em>III, and run on 1% agarose gel, contrast with 1KB ladder from NEB. <strong>B.</strong>&nbsp; Gel electrophoresis of pSB3T5-sucCD digested with <em>EcoR</em>I and <em>Hind</em>III, and run on 1% agarose gel, in the third well, there are two bands, the one located above is ringed DNAs are not be digested, sample loaded in this well is not cut completely.</p>
+<p style="text-align: justify;">Sinopec Guangzhou Petrochemical Company (&ldquo;Guangzhou Company&rdquo; for short) is one of the leading petrochemical enterprises in South China. The core business of Guangzhou Company covers refining&nbsp;products including solid plastics, such as polyethylene, polypropylene and polystyrene.</p>
+<p style="text-align: justify;">Jianfeng and Paul endorsed our project explaining that they welcome the technology development of bioplastic, even as a competitor to them. The mass convenience of plastic in food, clothes, car industry, etc. are too important and so far there is no other material that can replace plastic. And, currently the degradation of plastic is still too expensive for its price and its environmental impact. As a result, bioplastics constitute a possible solution to the growing demand of plastics.</p>
+     </blockquote>
+<div class="container"><button class="btn btn-info" type="button" data-toggle="collapse" data-target="#SINOPEC">Read more</button>
+<div id="SINOPEC" class="collapse">
+  <div class="col-sm-5">
+<p><strong>Understanding the recycling market of plastics </strong></p>
+<p style="text-align: justify;">Jianfeng and Paul have experience in the petroleum-based industry and knowledge of how the plastic market has change. We were aiming to understand the importance of different disposable scenarios and they gave us an insight of the recycling industry. They explained that after the reduction of oil prices in 2014, the recycling industry had retreated as making new plastic became much cheaper than recycling it. In addition, they mentioned that recycled plastics are slightly different and the manufacturers need to add other materials to improve the physical properties. These additives tend not be in the labels of the plastics making the recycling process more difficult in a further stage.</p>
+<p><strong>Implementations</strong></p>
+<p style="text-align: justify;">The recycling scenario would not be the best end-of-life for our plastics, in conjugation with the comments of Dr. Higson from NNFCC we believe that a combination of recycling and composting could be helpful in terms of the sustainability of our bioplastics. The results of the research in PHBV degradation and composability should be incorporated to the design of PHBV products.</p>
+</div>
+</div>
+</div>
+</div>
 </div>
 </div>
@@ Line 1,496: / Line 560: @@
+<!--11. Biome Bioplastics -->
-  <div class="container"><button class="btn btn-primary" type="button" data-toggle="collapse" data-target="#tittle53"><p><strong>Week 5 ---Week 7 Establish different <em>E. coli</em> trains </strong></p></button>
-        <div id="tittle53" class="collapse">
-<ul>
-<li>Transform recombinant plasmids to <em> coli</em> forming strains SC1 to SC11. (shown in Table 1)</li>
-</ul>
+    <div class="container"><button class="btn btn-warning" type="button" data-toggle="collapse" data-target="#tittle11"><h3><strong>&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;Biome Technologies&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;</strong></h3></button>
-<p style="text-align: center;"><strong>Table 1&nbsp; </strong>Basic information of engineered <em>E. coli</em> strain</p>
+        <div id="tittle11" class="collapse">
-<table style="margin-left: auto; margin-right: auto;" width="0">
-<tbody>
-<tr>
+   <div class="col-sm-5">
-<td width="180">
+<blockquote>
-<p>Bacterial strains:</p>
-</td>
+<p><strong>Paul Mines</strong>&ndash; CEO at Biome Technologies</p>
-<td width="378">
+<p><em>&ldquo;Plastics: wonder materials or existential threat&rdquo;</em></p>
-<p>Relevant information</p>
+<p style="text-align: justify;">Paul Mines began the lecture at The University of St Andrews by disclosing the issue of plastic pollution and the ubiquity of plastic due to its cheap and useful properties. Plastics have been around since 70 years ago, replacing other materials and expanding the plastic market. Then he continued discussing how bioplastic products can help to solve plastic pollution by offering plant-based and biodegradable alternatives to petroleum-derived plastics.</p>
-</td>
-</tr>
+     </blockquote>
-<tr>
-<td width="180">
-<p><em>Escherichia coli SC1</em></p>
-<p><em>Escherichia coli SC2</em></p>
-<p><em>Escherichia coli SC3</em></p>
-<p><em>Escherichia coli SC4</em></p>
-<p><em>Escherichia coli SC5</em></p>
-<p><em>Escherichia coli SC6</em></p>
-<p><em>Escherichia coli SC7</em></p>
-<p><em>Escherichia coli SC8</em></p>
-<p><em>Escherichia coli SC9</em></p>
-<p><em>Escherichia coli SC10</em></p>
-<p><em>Escherichia coli SC11</em></p>
-</td>
-<td width="378">
-<p><em>Escherichia coli BL21</em> containing pSB3T5-<em>sucAB</em></p>
-<p><em>Escherichia</em> coli BL21 containing pSB3T5-<em>sucCD</em></p>
-<p><em>Escherichia</em> coli BL21 containing pSB3T5-X</p>
-<p>BL21 containing pSB3T5-<em>sucAB</em> and pSB1C3-<em>PHA</em></p>
-<p>BL21 containing pSB3T5-<em>sucCD</em> and pSB1C3-<em>PHA</em></p>
-<p>BL21 containing pSB3T5-X and pSB1C3-<em>PHA</em></p>
-<p>BL21 containing pSB3T5-AB and empty pSB1C3</p>
-<p>BL21 containing pSB3T5-CD empty pSB1C3</p>
-<p>BL21 containing pSB3T5-X empty pSB1C3</p>
-<p>BL21 containing pSB3T5-<em>sucAB</em>-hypro and pSB1C3-<em>PHA</em></p>
-<p>BL21 containing pSB3T5-<em>sucCD</em>-hypor and pSB1C3-<em>PHA</em></p>
-</td>
-</tr>
-</tbody>
-</table>
-<p>&nbsp;</p>
-<ol start="2">
-<li>Pre-cultured strain SC1, SC2 and SC3 in LB medium</li>
-<li>Inoculate the pre-culture to M9 minimal medium with 1% glucose, 0.01M propionic acid and 10M IPTG.</li>
-<li>Optical density at 600nm was measured to plot the growth curve and growth rate was calculated using equation below.</li>
-</ol>
-<p style="text-align: center;"><strong>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; Table 2 </strong>Growth rate of <em>E. coli</em> strains SC1, SC2 and SC3</p>
-<table width="544">
-<tbody>
-<tr>
-<td style="text-align: center;" width="51">&nbsp;</td>
-<td style="text-align: center;" width="41">
-<p><strong>0 </strong></p>
-</td>
-<td style="text-align: center;" width="61">
-<p><strong>2 </strong></p>
-</td>
-<td style="text-align: center;" width="68">
-<p><strong>3 </strong></p>
-</td>
-<td style="text-align: center;" width="68">
-<p><strong>4 </strong></p>
-</td>
-<td style="text-align: center;" width="63">
-<p><strong>5 </strong></p>
-</td>
-<td style="text-align: center;">
-<p><strong>10 </strong></p>
-</td>
-<td style="text-align: center;">
-<p><strong>60 </strong></p>
-</td>
-<td style="text-align: center;">
-<p><strong>72 </strong></p>
-</td>
-</tr>
-<tr>
-<td style="text-align: center;" width="51">
-<p><strong>SC1 </strong></p>
-</td>
-<td style="text-align: center;" width="41">
-<p>0.18</p>
-</td>
-<td style="text-align: center;" width="61">
-<p>0.503667</p>
-</td>
-<td style="text-align: center;" width="68">
-<p>0.936667</p>
-</td>
-<td style="text-align: center;" width="68">
-<p>1.101</p>
-</td>
-<td style="text-align: center;" width="63">
-<p>1.195333</p>
-</td>
-<td style="text-align: center;">
-<p>1.379667</p>
-</td>
-<td style="text-align: center;">
-<p>1.3666</p>
-</td>
-<td style="text-align: center;">
-<p>1.3666</p>
-</td>
-</tr>
-<tr>
-<td style="text-align: center;" width="51">
-<p><strong>SC2 </strong></p>
-</td>
-<td style="text-align: center;" width="41">
-<p>0.18</p>
-</td>
-<td style="text-align: center;" width="61">
-<p>0.489333</p>
-</td>
-<td style="text-align: center;" width="68">
-<p>1.034</p>
-</td>
-<td style="text-align: center;" width="68">
-<p>1.238</p>
-</td>
-<td style="text-align: center;" width="63">
-<p>1.365</p>
-</td>
-<td style="text-align: center;">
-<p>1.440667</p>
-</td>
-<td style="text-align: center;">
-<p>1.36588</p>
-</td>
-<td style="text-align: center;">
-<p>1.36522</p>
-</td>
-</tr>
-<tr>
-<td style="text-align: center;" width="51">
-<p><strong>SC3 </strong></p>
-</td>
-<td style="text-align: center;" width="41">
-<p>0.28</p>
-</td>
-<td style="text-align: center;" width="61">
-<p>0.828</p>
-</td>
-<td style="text-align: center;" width="68">
-<p>1.497333</p>
-</td>
-<td style="text-align: center;" width="68">
-<p>1.618333</p>
-</td>
-<td style="text-align: center;" width="63">
-<p>1.755</p>
-</td>
-<td style="text-align: center;">
-<p>1.94</p>
-</td>
-<td style="text-align: center;">
-<p>1.9</p>
-</td>
-<td style="text-align: center;">&nbsp;</td>
-</tr>
-</tbody>
-</table>
-<p style="text-align: center;"><strong><img src="https://static.igem.org/mediawiki/2018/d/dc/T--Edinburgh_OG--Notebook_-_S3.png" width="500" height="315" /></strong></p>
-<p style="text-align: center;"><strong>Figure 3 </strong>Growth curves of strain SC1, strain SC2 and strain SC3 that harboured plasmid pSB3T5-sucAB, plasmid pSB3T5-sucCD and plasmid pSB3T5-X respectively. 1% glucose, 0.01M propionate and 10 &mu;M IPTG were added in the M9 medium.</p>
+<div class="container"><button class="btn btn-info" type="button" data-toggle="collapse" data-target="#Biome">Read more</button>
+<div id="Biome" class="collapse">
+  <div class="col-sm-5">
+<p><strong>Understanding the impacts of the process</strong></p>
+<p style="text-align: justify;">Regarding the carbon sequestration in bioplastics, the net carbon footprint is less and bioplastic can function as CO<sub>2</sub> storage, <strong>but</strong> however, it cannot help with the GHG problem in the world. Furthermore, he talked about the possible disposal scenarios such as composting, anaerobic digestion and incineration. Finally, he ended his presentation by emphasising on what needs to be developed to achieve the wide use of bioplastic. Some of his examples were: Plant science, biomass production, pre-treatment of the biomass, monomer treatment, and tests and scale up. Nevertheless, the market cannot go forward if the impact of bioplastics products to the environment and society is not understood.</p>
+<p style="text-align: justify;">The lecture reaffierded that &lsquo;Human Practices&rsquo; aspect is crucial in every project design and not just iGEM. Through our Human Practices journey, we found LCA tool which assessed the environmental impacts of a product and/or process, which is in line with what Paul Mines mentioned.</p>
+</div>
+</div>
 </div>
 </div>
-  <div class="container"><button class="btn btn-primary" type="button" data-toggle="collapse" data-target="#tittle54"><p><strong>Week 8 investigate the effect of <em>sucAB</em> and <em>sucCD</em> gene on growth and propionate adaption</strong></p></button>
-        <div id="tittle54" class="collapse">
-<ul>
-<li>Pre-culture six different strains to Investigate the effect of <em>sucAB</em> and <em>sucCD </em>gene on growth</li>
-<li>Measure the germinate multiple* (GM) of each strain, which represented the proliferation capacity of cells (shown in figure 4)</li>
-<p style="text-align: center;"><img src="https://static.igem.org/mediawiki/2018/2/23/T--Edinburgh_OG--Notebook_-_S4a.png" width="472" height="263" /><img src="https://static.igem.org/mediawiki/2018/e/ec/T--Edinburgh_OG--Notebook_-_S4b.png" width="474" height="260" /></p>
-<p><strong>Figure 4 </strong>Images of growth curves and Germinate Multiple -&nbsp;Stacked columns reflect growth rates of each strain under each case. Table at underneath the X-axis diaplays OD<sub>600</sub> measurement of six strains under serial propionate concentrations varying from 0 to 0.04 M at the interval of 0.01 M along with time line, time 0: immediately after strains are inoculated into M9 medium, 19: OD<sub>600</sub> was measured after cultivation of 19 hours, 24: OD<sub>600</sub> was measured after cultivation of 24 hours</p>
-<li>Determine the amount of propionic acid in the medium to investigate the effect of <em>sucAB</em> and <em>sucCD </em>gene on propionate uptake. Required standard curve was plotted during the pre-experimental. (shown in Figure 5)</li>
-<p style="text-align: center;"><strong><img style="display: block; margin-left: auto; margin-right: auto;" src="https://static.igem.org/mediawiki/2018/b/bc/T--Edinburgh_OG--Notebook_-_S5.png" width="447" height="272" />Figure 5</strong>&nbsp;Standard curve and the equation between the absorbance and propionate concentration.</p>
-<li>Obtain the amounts of propionic acid that utilized by <em> coli</em> strain SC7, SC8 and SC9.</li>
-</ul>
-<p><strong><img style="display: block; margin-left: auto; margin-right: auto;" src="https://static.igem.org/mediawiki/2018/3/32/T--Edinburgh_OG--Notebook_-_S6.png" width="461" height="278" /></strong></p>
-<p style="text-align: center;"><strong>Figure 6 </strong>The bar chart represented the amount of propionate taken by three different strains</p>
-<p>*GM defined as final cell concentration / inoculation cell concentration &nbsp;&nbsp;</p>
 </div>
 </div>
@@ Line 1,706: / Line 600: @@
-  <div class="container"><button class="btn btn-primary" type="button" data-toggle="collapse" data-target="#tittle55"><p><strong>Week 9 &ndash;Week 10 PHBV production and optimization </strong></p></button>
-        <div id="tittle55" class="collapse">
-<ul>
-<li>Dry the cells after cultivation of 60 hours.</li>
-<li>Nile red plate staining to confirm the production of PHBV</li>
-<p><strong><img style="display: block; margin-left: auto; margin-right: auto;" src="https://static.igem.org/mediawiki/2018/9/9a/T--Edinburgh_OG--Notebook_-_S7.png" width="529" height="280" /></strong></p>
-<p style="text-align: center;"><strong>Figure 7 </strong>Images of Nile red plate which are exposed to blue light or UV light;&nbsp;all six strains were spread on the plate for overnight culture&nbsp;</p>
-<li style="text-align: left;">Measure the cell dry weight (CDW).</li>
-<p style="text-align: center;"><strong><img src="https://static.igem.org/mediawiki/2018/6/61/T--Edinburgh_OG--Notebook_-_S8.png" width="482" height="237" /></strong></p>
-<p style="text-align: center;"><strong>Figure 8 </strong>Absolute cell dry weight of each strain against propionate concentrations,&nbsp;indicating the yield of each strains<strong> -</strong>&nbsp;It can be seen that when 0.03 M, all three strains have largest absolute dry cell weight, since cultured enough time, the same as PHB production, which corresponding to the largest absorption of propionate in 0.03M</p>
-<li>Change the concentration of IPTG. The effect of different IPTG concentration on cell growth and PHBV production were compared</li>
-<p><img style="display: block; margin-left: auto; margin-right: auto;" src="https://static.igem.org/mediawiki/2018/9/90/T--Edinburgh_OG--Notebook_-_S9.png" width="538" height="249" /></p>
-<p style="text-align: center;"><strong>Figure 9</strong>&nbsp;Growth of strain-SC7, SC8 and SC9 under different IPTG concentration, all the cases are cultured in the same condition. The line graph at left is the growth curve of strain-SC7 with IPTG concentration of 0, 0.05M and 0.1 M, the graph in the middle is from strain-SC8 and the line graph at right is strain-SC9.</p>
-<li>Measure the fluorescent intensity of three different strains using Plate Reader</li>
-</ul>
-<p><img style="display: block; margin-left: auto; margin-right: auto;" src="https://static.igem.org/mediawiki/2018/c/cd/T--Edinburgh_OG--Notebook_-_S10.png" width="555" height="282" /></p>
-<p style="text-align: center;"><strong>Figure 10 </strong>Bar chart of Fluorescent intensity - Cells were cultured with different concentrations of IPTG, fluorescent intensity was measured at cultivation of 24 hours and 48 hours</p>
-<p>Replace the T7 promoter with hybrid promoter, obtaining plasmids pSB3T5-sucAB-hypro and pSB3T5- sucCD-hypro</p>
-<p>&nbsp;</p>
-<p>&nbsp;</p>
-</div>
 </div>
@@ Line 1,741: / Line 605: @@
-<h2><strong>The Phasin and Hemolysin Secretion System (Owen)</strong></h2>
@@ Line 1,747: / Line 610: @@
-  <div class="container"><button class="btn btn-primary" type="button" data-toggle="collapse" data-target="#tittle61"><p><strong>Week 1 Design the strategy for plasmid construction&nbsp;</strong></p></button>
-        <div id="tittle61" class="collapse">
+    <div>
-<ul>
-<li>To investigate and optimized the&nbsp;level of Hemolysin transporter to PHB secretion, PCR strategy and digestion strategy were designed and ;utilised in plasmid construction.</li>
-<li>Order the primers</li>
-<p><img style="display: block; margin-left: auto; margin-right: auto;" src="https://static.igem.org/mediawiki/2018/5/50/T--Edinburgh_OG--Notebook_-_O1.png" /></span></p>
-<p style="text-align: center;"><strong>Figure 1&nbsp;</strong>Diagram of new&nbsp;Biobricks&nbsp;development -&nbsp;The development of Lac promoter-Phasin-HlyA&nbsp;without stop codon through PCR strategy. DNA from the Lac promoter-Phasin-HlyA&nbsp;original&nbsp;Biobrick&nbsp;was used as a template to remove the stop codon in the end of&nbsp;Phasin&nbsp;sequence. The PCR product was then digested with&nbsp;<em>Dpn</em>I&nbsp;(NEB) to remove the original DNA template then purified with&nbsp;QIAquick&nbsp;PCR Purification Kit (Qiagen), followed by self-ligation.</p>
-<p style="text-align: center;"><img src="https://static.igem.org/mediawiki/2018/7/73/T--Edinburgh_OG--Notebook_-_O2.png" /></p>
-<p style="text-align: center;"><strong>Figure 2&nbsp;</strong>Diagram&nbsp;of new&nbsp;Biobricks&nbsp;development<strong> -&nbsp;</strong>The development&nbsp;of&nbsp;pSB3T5-T7-hlyDB-Pro-phaP-hlyA. Several&nbsp;Biobricks&nbsp;were used in this process for assembly, these included T7 promoter, Lac Promoter-PhaP-HlyA,&nbsp;HlyA-tag+Secretion&nbsp;system and pSB3T5-I52001. The purple lines represent the location of enzyme digestion.&nbsp;HlyBD&nbsp;and T<span data-fontsize="12">7</span>&nbsp;promoter backbone was first obtained through digestion from their&nbsp;Biobricks&nbsp;then ligated together. The pSB1AK8 backbone of T<span data-fontsize="12">7</span>&nbsp;promoter-HlyBD&nbsp;then was replaced by digestion strategy to form T<span data-fontsize="12">7</span>&nbsp;promoter-HlyBD/pSB3T5. Lac promoter-Phasin-HlyA&nbsp;without stop codon were used as template to replace its promoter to J23100 promoter through PCT strategy. The parts of J23100 promoter-Phasin-HlyA&nbsp;and T<span data-fontsize="12">7</span>&nbsp;promoter-HlyBD/pSB3T5 vector were then ligated together to form T<span data-fontsize="12">7</span>&nbsp;promoter-HlyBD-J23100 promoter-Phasin-HlyA/pSB3T5.&nbsp;<span data-ccp-props="{&quot;335551550&quot;:6,&quot;335551620&quot;:6,&quot;335559685&quot;:567,&quot;335559740&quot;:360}">&nbsp;</span></p>
-<p style="text-align: center;"><span data-ccp-props="{&quot;335551550&quot;:6,&quot;335551620&quot;:6,&quot;335559685&quot;:567,&quot;335559740&quot;:360}"><img src="https://static.igem.org/mediawiki/2018/8/8f/T--Edinburgh_OG--Notebook_-_O3.png" width="546" height="293" /></span></p>
-<p style="text-align: center;"><span data-ccp-props="{&quot;335551550&quot;:6,&quot;335551620&quot;:6,&quot;335559685&quot;:567,&quot;335559740&quot;:360}"><span class="TextRun SCXW190868965" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW190868965"><strong>Figure 3</strong> Diagram of new&nbsp;</span><span class="SpellingError SCXW190868965">Biobricks</span><span class="NormalTextRun SCXW190868965">&nbsp;development.</span></span><span class="TextRun SCXW190868965" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW190868965">&nbsp;The development of&nbsp;</span></span><span class="TextRun SCXW190868965" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW190868965">pSB3T5-T7-hlyDB-</span></span><span class="TextRun SCXW190868965" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW190868965">phaR-</span></span><span class="TextRun SCXW190868965" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW190868965">phaP-hlyA</span></span><span class="EOP SCXW190868965" data-ccp-props="{&quot;134233117&quot;:true,&quot;134233118&quot;:true,&quot;335551550&quot;:6,&quot;335551620&quot;:6}">&nbsp;</span></span></p>
 <p>&nbsp;</p>
-<p><strong>Week&nbsp;</strong><strong>2</strong><strong>- Week&nbsp;</strong><strong>4</strong><strong>&nbsp;C</strong><strong>onstructs establishment&nbsp;</strong><strong>1</strong><span data-ccp-props="{&quot;134233117&quot;:true,&quot;134233118&quot;:true,&quot;335551550&quot;:6,&quot;335551620&quot;:6}">&nbsp;</span></p>
+</div>
-<ol style="font-weight: 400;">
-<li data-leveltext="%1." data-font="Calibri,DengXian" data-listid="2" aria-setsize="-1" data-aria-posinset="1" data-aria-level="1">Establish a new&nbsp;construct:&nbsp;DNA fragment pSB1AK8-T7-hlyBD&nbsp;(cloning strategy was shown&nbsp;in Figure 2)&nbsp;and transferred in&nbsp;<em>E. coli</em>&nbsp;BL21&nbsp;(DE3)</li>
-<li data-leveltext="%1." data-font="Calibri,DengXian" data-listid="2" aria-setsize="-1" data-aria-posinset="1" data-aria-level="1">Confirmation of successful&nbsp;pSB1AK8-T7-hlyBD&nbsp;plasmid&nbsp;with triple&nbsp;digestion</li>
+    <div>
-<li data-leveltext="%1." data-font="Calibri,DengXian" data-listid="2" aria-setsize="-1" data-aria-posinset="1" data-aria-level="1">Establish a new construct:&nbsp;DNA fragment pSB3T5-T7-hlyBD (cloning strategy was shown in Figure 2)&nbsp;and transferred in&nbsp;<em>E. coli</em>&nbsp;BL21&nbsp;(DE3)</li>
-<li data-leveltext="%1." data-font="Calibri,DengXian" data-listid="2" aria-setsize="-1" data-aria-posinset="1" data-aria-level="1">Confirmation of successful&nbsp;pSB3T5-T7-hlyBD&nbsp;plasmid&nbsp;with&nbsp;double&nbsp;digestion</li>
-</ol>
-<p style="text-align: center;"><span class="TextRun SCXW118191532" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW118191532"><strong><img src="https://static.igem.org/mediawiki/2018/2/2c/T--Edinburgh_OG--Notebook_-_O4i.png" /><img src="https://static.igem.org/mediawiki/2018/0/01/T--Edinburgh_OG--Notebook_-_O4ii.png" width="283" height="244" /></strong></span></span></p>
-<p style="text-align: center;"><span class="TextRun SCXW118191532" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW118191532"><strong>Figure 4</strong> Agarose gel electrophoresis of restriction enzyme-digested&nbsp;</span><span class="SpellingError SCXW118191532">Biobricks</span><span class="NormalTextRun SCXW118191532">.</span></span><span class="TextRun SCXW118191532" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW118191532">&nbsp;</span></span><span class="TextRun SCXW118191532" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW118191532">A</span></span><span class="TextRun SCXW118191532" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW118191532">.&nbsp;</span></span><span class="TextRun SCXW118191532" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW118191532">pSB1AK8-T7-hlyBD&nbsp;</span></span><span class="TextRun SCXW118191532" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW118191532">plasmid (lane 1: 1 kb DNA ladder; 2: undigested plasmid; 3:&nbsp;</span></span><span class="TextRun SCXW118191532" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW118191532">EcoR</span></span><span class="TextRun SCXW118191532" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW118191532">1 digestion; 4:&nbsp;</span></span><span class="TextRun SCXW118191532" lang="EN-US" xml:lang="EN-US"><span class="SpellingError SCXW118191532">Hind</span></span><span class="TextRun SCXW118191532" lang="EN-US" xml:lang="EN-US"><span class="SpellingError SCXW118191532">III</span><span class="NormalTextRun SCXW118191532">&nbsp;digestion; 5:&nbsp;</span></span><span class="TextRun SCXW118191532" lang="EN-US" xml:lang="EN-US"><span class="SpellingError SCXW118191532">Pst</span></span><span class="TextRun SCXW118191532" lang="EN-US" xml:lang="EN-US"><span class="SpellingError SCXW118191532">I</span></span><span class="TextRun SCXW118191532" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW118191532">&nbsp;digestion; 6:&nbsp;</span></span><span class="TextRun SCXW118191532" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW118191532">EcoR</span></span><span class="TextRun SCXW118191532" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW118191532">1,&nbsp;</span></span><span class="TextRun SCXW118191532" lang="EN-US" xml:lang="EN-US"><span class="SpellingError SCXW118191532">Hind</span></span><span class="TextRun SCXW118191532" lang="EN-US" xml:lang="EN-US"><span class="SpellingError SCXW118191532">III</span><span class="NormalTextRun SCXW118191532">&nbsp;and&nbsp;</span></span><span class="TextRun SCXW118191532" lang="EN-US" xml:lang="EN-US"><span class="SpellingError SCXW118191532">Pst</span></span><span class="TextRun SCXW118191532" lang="EN-US" xml:lang="EN-US"><span class="SpellingError SCXW118191532">I</span></span><span class="TextRun SCXW118191532" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW118191532">&nbsp;triple digestion).&nbsp;</span></span><span class="TextRun SCXW118191532" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW118191532">B</span></span><span class="TextRun SCXW118191532" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW118191532">.&nbsp;</span></span><span class="TextRun SCXW118191532" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW118191532">pSB3T5-T7-hlyBD&nbsp;</span></span><span class="TextRun SCXW118191532" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW118191532">(lane 0 &amp; 1: 1 kb DNA ladder; 2: undigested plasmid; 3:&nbsp;</span></span><span class="TextRun SCXW118191532" lang="EN-US" xml:lang="EN-US"><span class="SpellingError SCXW118191532">Spe</span></span><span class="TextRun SCXW118191532" lang="EN-US" xml:lang="EN-US"><span class="SpellingError SCXW118191532">I</span></span><span class="TextRun SCXW118191532" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW118191532">&nbsp;digestion; 4:&nbsp;</span></span><span class="TextRun SCXW118191532" lang="EN-US" xml:lang="EN-US"><span class="SpellingError SCXW118191532">Pst</span></span><span class="TextRun SCXW118191532" lang="EN-US" xml:lang="EN-US"><span class="SpellingError SCXW118191532">I</span></span><span class="TextRun SCXW118191532" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW118191532">&nbsp;digestion; 5:&nbsp;</span></span><span class="TextRun SCXW118191532" lang="EN-US" xml:lang="EN-US"><span class="SpellingError SCXW118191532">Spe</span></span><span class="TextRun SCXW118191532" lang="EN-US" xml:lang="EN-US"><span class="SpellingError SCXW118191532">I</span></span><span class="TextRun SCXW118191532" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW118191532">&nbsp;and&nbsp;</span></span><span class="TextRun SCXW118191532" lang="EN-US" xml:lang="EN-US"><span class="SpellingError SCXW118191532">Pst</span></span><span class="TextRun SCXW118191532" lang="EN-US" xml:lang="EN-US"><span class="SpellingError SCXW118191532">I</span></span><span class="TextRun SCXW118191532" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW118191532">&nbsp;double digestion)</span></span></p>
 <p>&nbsp;</p>
 </div>
+    <div>
+<p>&nbsp;</p>
 </div>
+    <div>
-  <div class="container"><button class="btn btn-primary" type="button" data-toggle="collapse" data-target="#tittle62"><p><strong>Week</strong><strong>&nbsp;4</strong><strong>&nbsp;&ndash; Week&nbsp;</strong><strong>6</strong><strong>.&nbsp;&nbsp;</strong><strong>Constructs establishment</strong><strong>&nbsp;2&nbsp;</strong><span data-ccp-props="{&quot;335551550&quot;:6,&quot;335551620&quot;:6}">&nbsp;</span></p></button>
-        <div id="tittle62" class="collapse">
-<ol>
-<li data-leveltext="%1." data-font="Calibri,DengXian" data-listid="9" aria-setsize="-1" data-aria-posinset="1" data-aria-level="1">Stop codon removal from the original&nbsp;Biobrick&nbsp;(Lac promoter-Phasin-Hemolysin A/&nbsp;pSB1C3, LPH/pSB1C3)&nbsp;with PCR strategy&nbsp;as shown in Figure 1 and transferred in&nbsp;<em>E. coli</em>&nbsp;BL21(DE3)</li>
-<li data-leveltext="%1." data-font="Calibri,DengXian" data-listid="9" aria-setsize="-1" data-aria-posinset="1" data-aria-level="1">Confirmation&nbsp;of&nbsp;with&nbsp;Stop codon removed LPH/pSB1C3 via&nbsp;<em>EcoR</em>I&nbsp;and&nbsp;<em>Hind</em>III&nbsp;double digestion, which the&nbsp;<em>Hind</em>III&nbsp;restriction enzyme was not present in the original&nbsp;Biobrick&nbsp;and introduced through the PCR amplification</li>
-<li data-leveltext="%1." data-font="Calibri,DengXian" data-listid="9" aria-setsize="-1" data-aria-posinset="1" data-aria-level="1">Establishment of two&nbsp;new constructs:&nbsp;pSB1C3-T7-hlyBD-Pro-phaP-hlyA and pSB1C3-T7-hlyBD-phaR-phaP-hlyA&nbsp;and transferred in&nbsp;<em>E. coli</em>&nbsp;BL21(DE3)</li>
-<li data-leveltext="%1." data-font="Calibri,DengXian" data-listid="9" aria-setsize="-1" data-aria-posinset="1" data-aria-level="1">Confirmation of&nbsp;pSB1C3-T7-hlyBD-JPH&nbsp;and&nbsp;pSB1C3-RPH&nbsp;with&nbsp;<em>EcoR</em>I&nbsp;and&nbsp;<em>Pst</em>I&nbsp;double&nbsp;digestion</li>
-</ol>
 <p>&nbsp;</p>
-<p style="text-align: left;"><strong>Week&nbsp;7&nbsp;&ndash;&nbsp;Week&nbsp;9&nbsp;Cell culture&nbsp;for growth study&nbsp;&nbsp;</strong></p>
-<ol>
-<li data-leveltext="%1." data-font="Calibri" data-listid="6" aria-setsize="-1" data-aria-posinset="1" data-aria-level="1">Culture the&nbsp;<em>E. coli</em>&nbsp;BL21&nbsp;(DE3), which&nbsp;harbouring&nbsp;the following plasmid(s)&nbsp;in M9 medium that contained 3% glucose&nbsp;with corresponding antibiotic(s) concentration, and&nbsp;assessed&nbsp;their optical density&nbsp;in&nbsp;different&nbsp;time points
-<ol style="list-style-type: lower-alpha;">
-<li data-leveltext="%1." data-font="Calibri" data-listid="6" aria-setsize="-1" data-aria-posinset="1" data-aria-level="1">pSB1C3,&nbsp;LPH,&nbsp;LPH&nbsp;(without stop codon),&nbsp;pSB1C3&nbsp;(Red fluorescent&nbsp;protein+),&nbsp;pSB1C3&nbsp;(RFP-),&nbsp;pSB1C3, PHA operon, PHA operon +&nbsp;pSB1C3&nbsp;(0, 3, 21, 25, 47, 51 and 71hours);&nbsp;&nbsp;<span data-ccp-props="{&quot;134233279&quot;:true,&quot;335551550&quot;:6,&quot;335551620&quot;:6}">&nbsp;</span></li>
-<li data-leveltext="%1." data-font="Calibri" data-listid="6" aria-setsize="-1" data-aria-posinset="1" data-aria-level="1">T7 promoter-HlyBD&nbsp;-&nbsp;JPH (0, 4, 7.5,&nbsp;24, 28 and 94 hours); T7 promoter-HlyBD&nbsp;-&nbsp;JPH&nbsp;+ PHA operon (0, 3, 20, 24 and 90 hours); in the case of IPTG induction, pSB3T5 (RFP+) and T7 promoter-HlyBD&nbsp;(0, 3.5, 4.5 and 72 hours), T7 promoter-HlyBD&nbsp;-&nbsp;JPH&nbsp;(0, 3.5, 5.5 and 71.5 hours), T7 promoter-HlyBD&nbsp;-&nbsp;JPH&nbsp;+ PHA operon (0, 2.5, 19.5, 24.5, 42 and 47 hours),&nbsp;LPH&nbsp;(0, 3.5, 5.5, 71.5, 75.5, 78, 95, 100 and 117 hours), and&nbsp;LPH&nbsp;(without stop codon) (0, 3.5, 5.5, 17, 23 and 71.5 hours)</li>
-<li data-leveltext="%1." data-font="Calibri" data-listid="6" aria-setsize="-1" data-aria-posinset="1" data-aria-level="1"><em>phaCAB</em>&nbsp;operon&nbsp;+&nbsp;pSB1C3&nbsp;incubated in 50ml culture (250ml Flask) were measured at 0, 18, 24.5, 39.5, 43.5 and 63.5 hours)</li>
-</ol>
-</ol>
-<p><span data-ccp-props="{&quot;335551550&quot;:6,&quot;335551620&quot;:6}">&nbsp;<img style="float: left;" src="https://static.igem.org/mediawiki/2018/8/86/T--Edinburgh_OG--Notebook_-_O5i.png" width="595" height="277" /><img style="float: left;" src="https://static.igem.org/mediawiki/2018/c/c5/T--Edinburgh_OG--Notebook_-_O5ii.png" /></span></p>
-<p>&nbsp;&nbsp;&nbsp;</p>
-<p style="text-align: center;">&nbsp;&nbsp;&nbsp;<strong>Figure 5 </strong>Agarose gel electrophoresis of&nbsp;Phasin-HlyA&nbsp;products.&nbsp;A. PCR product of Lac promoter-Phasin-HlyA&nbsp;with stop codon removal.&nbsp;B. Enzyme digestion of Lac promoter-Phasin-HlyA&nbsp;PCR product for stop codon removal (lane 1: 1 kb DNA ladder; 2:&nbsp;<em>EcoR</em>I&nbsp;and&nbsp;<em>Hind</em>III&nbsp;&nbsp; double digestion) with the label of&nbsp;HlyA&nbsp;+ pSB1C3 Backbone and&nbsp;Phasin.&nbsp;C. PCR product of&nbsp;pSB1C3-Lac promoter-Phasin-HlyA&nbsp;(stop codon -) with J23100 promoter forward and reverse primers 1;&nbsp;D. PCR product of&nbsp;pSB1C3-Lac promoter-Phasin-HlyA&nbsp;(stop codon -) with J23100 promoter forward and reverse primers 2;&nbsp;E. Enzyme digestion of constructed T7 promoter-HlyBD-JPH plasmid andT7 promoter-HlyBD-RPH plasmid (lane 1 &amp; 4: 1&nbsp;kd&nbsp;DNA ladder; 2-3:&nbsp;<em>EcoR</em>I&nbsp;and&nbsp;<em>Pst</em>I&nbsp;double digestion for T7 promoter-&nbsp;HlyBD-JPH plasmid; 5-6&nbsp;<em>EcoR</em>I&nbsp;and&nbsp;<em>Pst</em>I&nbsp;double digestion for&nbsp;T7 promoter-HlyBD-RPH plasmid).&nbsp;</p>
-<p><img style="display: block; margin-left: auto; margin-right: auto;" src="https://static.igem.org/mediawiki/2018/a/af/T--Edinburgh_OG--Notebook_-_O6.png" /></p>
-<p><strong>Week 9&nbsp;</strong><strong>Determination of&nbsp;</strong><strong>PHA production&nbsp;</strong><span data-ccp-props="{&quot;335551550&quot;:6,&quot;335551620&quot;:6}">&nbsp;</span></p>
-<ol style="font-weight: 400;">
-<li data-leveltext="%1." data-font="Calibri" data-listid="7" aria-setsize="-1" data-aria-posinset="1" data-aria-level="1">Nile red plate staining<span data-ccp-props="{&quot;134233279&quot;:true,&quot;335551550&quot;:6,&quot;335551620&quot;:6}">&nbsp; &nbsp;</span></li>
-</ol>
-<p><span data-ccp-props="{&quot;134233279&quot;:true,&quot;335551550&quot;:6,&quot;335551620&quot;:6}"><img style="display: block; margin-left: auto; margin-right: auto;" src="https://static.igem.org/mediawiki/2018/d/d6/T--Edinburgh_OG--Notebook_-_O7.png" /></span></p>
-<p style="text-align: center;"><span data-ccp-props="{&quot;134233279&quot;:true,&quot;335551550&quot;:6,&quot;335551620&quot;:6}"><span class="TextRun SCXW224013829" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW224013829"><strong>Figure 7</strong> The growth study of&nbsp;</span></span><span class="TextRun SCXW224013829" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW224013829">E.</span></span><span class="TextRun SCXW224013829" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW224013829">&nbsp;</span></span><span class="TextRun SCXW224013829" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW224013829">coli</span></span><span class="TextRun SCXW224013829" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW224013829">&nbsp;Bl21</span></span><span class="TextRun SCXW224013829" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW224013829">&nbsp;</span></span><span class="TextRun SCXW224013829" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW224013829">(DE3) strain with constructed&nbsp;</span><span class="SpellingError SCXW224013829">HlyBD-Phasin-HlyA</span><span class="NormalTextRun SCXW224013829">&nbsp;plasmid with and without IPTG (Triplicate).</span></span><span class="TextRun SCXW224013829" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW224013829">&nbsp;The results are represented as the mean&nbsp;</span></span><span class="TextRun SCXW224013829" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW224013829">OD</span></span><span class="TextRun SCXW224013829" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW224013829" data-fontsize="11">600</span></span><span class="TextRun SCXW224013829" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW224013829">&nbsp;&plusmn; S.E.M.</span></span></span></p>
-<p style="text-align: center;"><span data-ccp-props="{&quot;134233279&quot;:true,&quot;335551550&quot;:6,&quot;335551620&quot;:6}"><span class="TextRun SCXW224013829" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW224013829"><img src="https://static.igem.org/mediawiki/2018/4/4f/T--Edinburgh_OG--Notebook_-_O8.png" /></span></span></span></p>
-<p style="text-align: center;"><span data-ccp-props="{&quot;134233279&quot;:true,&quot;335551550&quot;:6,&quot;335551620&quot;:6}"><span class="TextRun SCXW224013829" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW224013829"><span class="TextRun SCXW133904053" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW133904053"><strong>Figure 8</strong> The growth study of PHA operon -</span></span><span class="TextRun SCXW133904053" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW133904053">&nbsp;The results are represented as the mean&nbsp;</span></span><span class="TextRun SCXW133904053" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW133904053">OD</span></span><span class="TextRun SCXW133904053" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW133904053" data-fontsize="11">600</span></span><span class="TextRun SCXW133904053" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW133904053">&nbsp;&plusmn; S.E.M.&nbsp;</span></span><strong><span class="TextRun SCXW133904053" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW133904053">A.</span></span></strong><span class="TextRun SCXW133904053" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW133904053">&nbsp;The growth study of&nbsp;</span></span><span class="TextRun SCXW133904053" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW133904053">E.</span></span><span class="TextRun SCXW133904053" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW133904053">&nbsp;</span></span><span class="TextRun SCXW133904053" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW133904053">coli</span></span><span class="TextRun SCXW133904053" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW133904053">&nbsp;BL21</span></span><span class="TextRun SCXW133904053" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW133904053">&nbsp;</span></span><span class="TextRun SCXW133904053" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW133904053">(DE3) strain with pSB1C3, PHA operon,&nbsp;</span></span><span class="TextRun SCXW133904053" lang="EN-US" xml:lang="EN-US"><span class="SpellingError SCXW133904053">phaCAB</span></span><span class="TextRun SCXW133904053" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW133904053">&nbsp;operon+</span></span><span class="TextRun SCXW133904053" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW133904053">&nbsp;</span></span><span class="TextRun SCXW133904053" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW133904053">pSB3T5</span></span><span class="TextRun SCXW133904053" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW133904053">&nbsp;</span></span><span class="TextRun SCXW133904053" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW133904053">(RFP-) in 50ml tube (performed in triplicate).&nbsp;</span></span><strong><span class="TextRun SCXW133904053" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW133904053">B.</span></span></strong><span class="TextRun SCXW133904053" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW133904053">&nbsp;The growth study of&nbsp;</span></span><span class="TextRun SCXW133904053" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW133904053">E.</span></span><span class="TextRun SCXW133904053" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW133904053">&nbsp;</span></span><span class="TextRun SCXW133904053" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW133904053">coli</span></span><span class="TextRun SCXW133904053" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW133904053">&nbsp;BL21</span></span><span class="TextRun SCXW133904053" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW133904053">&nbsp;</span></span><span class="TextRun SCXW133904053" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW133904053">(DE3) strain with&nbsp;</span></span><span class="TextRun SCXW133904053" lang="EN-US" xml:lang="EN-US"><span class="SpellingError SCXW133904053">phaCAB</span></span><span class="TextRun SCXW133904053" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW133904053">&nbsp;operon+pSB3T5</span></span><span class="TextRun SCXW133904053" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW133904053">&nbsp;</span></span><span class="TextRun SCXW133904053" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW133904053">(RFP-) in 250ml Flask.&nbsp;</span></span><span class="EOP SCXW133904053" data-ccp-props="{&quot;335551550&quot;:6,&quot;335551620&quot;:6,&quot;335559685&quot;:567}">&nbsp;</span></span></span></span></p>
-<p style="text-align: center;"><span data-ccp-props="{&quot;134233279&quot;:true,&quot;335551550&quot;:6,&quot;335551620&quot;:6}"><span class="TextRun SCXW224013829" lang="EN-US" xml:lang="EN-US"><span class="NormalTextRun SCXW224013829"><span class="EOP SCXW133904053" data-ccp-props="{&quot;335551550&quot;:6,&quot;335551620&quot;:6,&quot;335559685&quot;:567}"><img src="https://static.igem.org/mediawiki/2018/3/36/T--Edinburgh_OG--Notebook_-_O9.png" /></span></span></span></span></p>
-<p style="text-align: center;"><strong>Figure 9</strong> The growth study of&nbsp;<em>E.</em><em>&nbsp;</em><em>coli</em>&nbsp;Bl21&nbsp;(DE3) strain with&nbsp;pSB3T5-T7-hlyDB-phaP-hlyA and&nbsp;<em>phaCAB</em>&nbsp;operon&nbsp;with and without IPTG (Triplicate). The results are represented as the mean&nbsp;OD<span data-fontsize="11">600</span><span data-fontsize="11">&nbsp;</span>&plusmn; S.E.M.&nbsp;&nbsp;&nbsp;<span data-ccp-props="{&quot;335551550&quot;:6,&quot;335551620&quot;:6,&quot;335559685&quot;:567}">&nbsp;</span></p>
-<p><strong>Week&nbsp;</strong><strong>9</strong><strong>&nbsp;&ndash;&nbsp;</strong><strong>W</strong><strong>eek&nbsp;</strong><strong>10</strong><strong>&nbsp;</strong><strong>C</strong><strong>ell culture&nbsp;</strong><strong>for PHA production</strong><span data-ccp-props="{&quot;335551550&quot;:6,&quot;335551620&quot;:6}">&nbsp;</span></p>
-<ol>
-<li data-leveltext="%1." data-font="Calibri" data-listid="10" aria-setsize="-1" data-aria-posinset="1" data-aria-level="1">Culture the&nbsp;<em>E. coli</em>&nbsp;BL21&nbsp;(DE3)&nbsp;to investigate PHA production in different condition under&nbsp;M9 medium that contained 3% glucose with corresponding antibiotic(s) concentration</li>
-<li data-leveltext="%1." data-font="Calibri" data-listid="10" aria-setsize="-1" data-aria-posinset="1" data-aria-level="1">Measure fluorescent intensity of cultures to provide real-time information of PHA production. (semi-quantitative Nile red measurement)<span data-ccp-props="{&quot;134233279&quot;:true,&quot;335551550&quot;:6,&quot;335551620&quot;:6}">&nbsp;</span></li>
-</ol>
-<p style="text-align: center;"><span data-ccp-props="{&quot;335551550&quot;:6,&quot;335551620&quot;:6,&quot;335559685&quot;:360}">&nbsp;</span><strong>Table&nbsp;</strong><strong>1</strong><strong>&nbsp;</strong>PHB production of recombinant&nbsp;<em>E.</em><em>&nbsp;</em><em>coli</em>&nbsp;BL21&nbsp;(DE3) strain with PHA operon from 72 hours bacterial culture in M9 medium with 0.3% glucose (performed in singular).&nbsp;The results of&nbsp;OD600&nbsp;are represented as the mean OD600&nbsp;&plusmn;&nbsp;S.E.M.&nbsp;</p>
-<table>
-<tbody>
-<tr>
-<td data-celllook="0">
-<p><span data-ccp-props="{&quot;335551550&quot;:6,&quot;335551620&quot;:6}">&nbsp;</span></p>
-</td>
-<td data-celllook="0">
-<p><strong>Dry cell mass (g)&nbsp;&nbsp;</strong></p>
-</td>
-<td data-celllook="0">
-<p><strong>PHB production (g)&nbsp;</strong></p>
-</td>
-<td data-celllook="0">
-<p><strong>Melting Temperature (&deg;C)&nbsp;</strong></p>
-</td>
-<td data-celllook="0">
-<p><strong>OD600&nbsp;</strong></p>
-</td>
-</tr>
-<tr>
-<td data-celllook="0">
-<p>1 litre flask incubation (200ml culture sample)<span data-ccp-props="{&quot;335551550&quot;:6,&quot;335551620&quot;:6}">&nbsp;</span></p>
-</td>
-<td data-celllook="0">
-<p style="text-align: center;">0.59&nbsp;</p>
-</td>
-<td style="text-align: center;" data-celllook="0">
-<p>0.008&nbsp;</p>
-</td>
-<td style="text-align: center;" data-celllook="0">
-<p>165-170&nbsp;</p>
-</td>
-<td data-celllook="0">
-<p style="text-align: center;">2.244 &plusmn;&nbsp;</p>
-<p style="text-align: center;">0.014<strong>&nbsp;</strong></p>
-</td>
-</tr>
-<tr>
-<td data-celllook="0">
-<p>500ml flask incubation (100ml culture sample)<span data-ccp-props="{&quot;335551550&quot;:6,&quot;335551620&quot;:6}">&nbsp;</span></p>
-</td>
-<td data-celllook="0">
-<p style="text-align: center;">0.285&nbsp;</p>
-</td>
-<td style="text-align: center;" data-celllook="0">
-<p>0.001&nbsp;&nbsp;&nbsp;</p>
-</td>
-<td style="text-align: center;" data-celllook="0">
-<p>168-175&nbsp;</p>
-</td>
-<td data-celllook="0">
-<p style="text-align: center;">1.629 &plusmn;&nbsp;</p>
-<p style="text-align: center;">0.035&nbsp;</p>
-</td>
-</tr>
-<tr>
-<td data-celllook="0">
-<p>250ml flask incubation (50ml culture sample)<span data-ccp-props="{&quot;335551550&quot;:6,&quot;335551620&quot;:6}">&nbsp;</span></p>
-</td>
-<td style="text-align: center;" data-celllook="0">
-<p>0.17&nbsp;</p>
-</td>
-<td style="text-align: center;" data-celllook="0">
-<p>Not measurable&nbsp;&nbsp;&nbsp;</p>
-</td>
-<td style="text-align: center;" data-celllook="0">
-<p>&nbsp;160-169&nbsp;</p>
-</td>
-<td style="text-align: center;" data-celllook="0">
-<p>1.913 &plusmn;&nbsp;</p>
-<p>0.013&nbsp;</p>
-</td>
-</tr>
-</tbody>
-</table>
-<p>&nbsp;<br /><span data-ccp-props="{}">&nbsp;</span></p>
-<p><strong>Table&nbsp;2</strong> The PHB production of&nbsp;<em>E.</em><em>&nbsp;</em><em>coli</em>&nbsp;BL21&nbsp;(DE3) strain (PHA operon + pSB3T5) cell culture for 62 hours in M9 medium with 3% glucose (performed in duplicate)&nbsp;&nbsp;</p>
-<table>
-<tbody>
-<tr>
-<td colspan="1" rowspan="2" data-celllook="0">
-<p><span data-ccp-props="{&quot;335551550&quot;:2,&quot;335551620&quot;:2}">&nbsp;</span></p>
-</td>
-<td colspan="3" data-celllook="0">
-<p>Extracellular fraction<span data-ccp-props="{&quot;335551550&quot;:2,&quot;335551620&quot;:2}">&nbsp;</span></p>
-</td>
-<td colspan="2" data-celllook="0">
-<p>Intracellular&nbsp;fraction<span data-ccp-props="{&quot;335551550&quot;:2,&quot;335551620&quot;:2}">&nbsp;</span></p>
-</td>
-<td colspan="1" rowspan="2" data-celllook="0">
-<p>PHB Production<span data-ccp-props="{&quot;335551550&quot;:2,&quot;335551620&quot;:2}">&nbsp;</span></p>
-</td>
-</tr>
-<tr>
-<td data-celllook="0">
-<p>Amount of PHB in<span data-ccp-props="{&quot;335551550&quot;:2,&quot;335551620&quot;:2}">&nbsp;</span></p>
-</td>
-<td data-celllook="0">
-<p>CaCl<span data-fontsize="11">2</span>&nbsp;added<span data-ccp-props="{&quot;335551550&quot;:2,&quot;335551620&quot;:2}">&nbsp;</span></p>
-</td>
-<td data-celllook="0">
-<p>Corrected amount of PHB<span data-ccp-props="{&quot;335551550&quot;:2,&quot;335551620&quot;:2}">&nbsp;</span></p>
-</td>
-<td data-celllook="0">
-<p>Amount of PHB<span data-ccp-props="{&quot;335551550&quot;:2,&quot;335551620&quot;:2}">&nbsp;</span></p>
-</td>
-<td data-celllook="0">
-<p>Melting Temperature<span data-ccp-props="{&quot;335551550&quot;:2,&quot;335551620&quot;:2}">&nbsp;</span></p>
-</td>
-</tr>
-<tr>
-<td data-celllook="0">
-<p>50ml culture sample X 2<span data-ccp-props="{&quot;335551550&quot;:2,&quot;335551620&quot;:2}">&nbsp;</span></p>
-</td>
-<td data-celllook="0">
-<p>0.016&plusmn;0g<span data-ccp-props="{&quot;335551550&quot;:2,&quot;335551620&quot;:2}">&nbsp;</span></p>
-</td>
-<td data-celllook="0">
-<p>0.110g<span data-ccp-props="{&quot;335551550&quot;:2,&quot;335551620&quot;:2}">&nbsp;</span></p>
-</td>
-<td data-celllook="0">
-<p>0g<span data-ccp-props="{&quot;335551550&quot;:2,&quot;335551620&quot;:2}">&nbsp;</span></p>
-</td>
-<td data-celllook="0">
-<p>0.011&plusmn;0g<span data-ccp-props="{&quot;335551550&quot;:2,&quot;335551620&quot;:2}">&nbsp;</span></p>
-</td>
-<td data-celllook="0">
-<p>165-170&deg;C<span data-ccp-props="{&quot;335551550&quot;:2,&quot;335551620&quot;:2}">&nbsp;</span></p>
-</td>
-<td data-celllook="0">
-<p>0.11g/L<span data-ccp-props="{&quot;335551550&quot;:2,&quot;335551620&quot;:2}">&nbsp;</span></p>
-</td>
-</tr>
-<tr>
-<td data-celllook="0">
-<p>50 ml culture sample<span data-ccp-props="{&quot;335551550&quot;:2,&quot;335551620&quot;:2}">&nbsp;</span></p>
-</td>
-<td data-celllook="0">
-<p>0.0085&plusmn;0g<span data-ccp-props="{&quot;335551550&quot;:2,&quot;335551620&quot;:2}">&nbsp;</span></p>
-</td>
-<td data-celllook="0">
-<p>0.055g<span data-ccp-props="{&quot;335551550&quot;:2,&quot;335551620&quot;:2}">&nbsp;</span></p>
-</td>
-<td data-celllook="0">
-<p>0g<span data-ccp-props="{&quot;335551550&quot;:2,&quot;335551620&quot;:2}">&nbsp;</span></p>
-</td>
-<td data-celllook="0">
-<p>0.006&plusmn;0g<span data-ccp-props="{&quot;335551550&quot;:2,&quot;335551620&quot;:2}">&nbsp;</span></p>
-</td>
-<td data-celllook="0">
-<p>168-175&deg;C<span data-ccp-props="{&quot;335551550&quot;:2,&quot;335551620&quot;:2}">&nbsp;</span></p>
-</td>
-<td data-celllook="0">
-<p>0.12g/L<span data-ccp-props="{&quot;335551550&quot;:2,&quot;335551620&quot;:2}">&nbsp;</span></p>
-</td>
-</tr>
-</tbody>
-</table>
-<p style="text-align: center;"><span data-ccp-props="{&quot;335551550&quot;:6,&quot;335551620&quot;:6}"><img src="https://static.igem.org/mediawiki/2018/a/a9/T--Edinburgh_OG--Notebook_-_O10.png" />&nbsp;</span></p>
-<p style="text-align: center;"><strong>Figure 10 </strong>Extracted product via PHB extraction protocol -&nbsp;<strong>A.</strong> Extracted product from intracellular fragment. <strong>B.</strong> Extracted product from&nbsp;extracellular&nbsp;fragment.&nbsp;&nbsp;&nbsp;&nbsp;</p>
-<p>&nbsp;<img style="display: block; margin-left: auto; margin-right: auto;" src="https://static.igem.org/mediawiki/2018/4/4a/T--Edinburgh_OG--Notebook_-_O11.png" width="291" height="270" /></p>
-<p style="text-align: center;"><strong>Figure 1</strong><strong>1</strong><strong>&nbsp;</strong>Fluorescence intensity detection of&nbsp;<em>E.</em><em>&nbsp;</em><em>coli</em>&nbsp;BL21(DE3) strain with&nbsp;pSB1C3-T7-hlyBD-JPH and<em>&nbsp;phaCAB</em>&nbsp;operon under Nile red stain (performed in triplicate) -&nbsp;Results are represented as the mean fluorescent strength &plusmn; S.E.M. measured at 520 nm excitation and 590 nm emission wavelengths in 24 and 48 hours.&nbsp;Cultured in M9 medium with 3% glucose (performed in duplicate)</p>
-</div>
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-</div>
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Difference between revisions of "Team:Edinburgh OG/Human Practices"

Latest revision as of 03:57, 18 October 2018

Integrated Human Practices

Dialogue with

Dialogue with

Dialogue with

Dialogue with

MSc students in Tools of Synthetic Biology class 2017/2018

Commercial Links

The National Non-Food Crops Centre (NNFCC)

Life Cycle Assessment (LCA)

Cambridge Consultants

iGEM Northern UK Meet-up @ U St Andrews

Scotch Whisky Association

Visit to Local Whisky Distillery in Edinburgh

BLUEPHA Technologies

SINOPEC

Biome Technologies

MSc students in Tools of Synthetic Biology class
2017/2018