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<hr class="line"> | <hr class="line"> | ||
− | <img src="https://static.igem.org/mediawiki/2019/a/ac/T--Marburg--logo.svg" class="logo" alt="Syntex Logo"> | + | <img src="https://static.igem.org/mediawiki/2019/a/ac/T--Marburg--logo.svg" |
+ | class="logo" | ||
+ | alt="Syntex Logo"> | ||
</div> | </div> | ||
<div style="margin-top: 10vh;"> | <div style="margin-top: 10vh;"> | ||
<section class="section"> | <section class="section"> | ||
− | |||
<p style="text-align: justify;"> | <p style="text-align: justify;"> | ||
− | Through this iGEM year we interacted with | + | Through this iGEM year we interacted with many iGEM Teams all around the world. It was a pleasure working |
− | with all the different teams and learn from each other. We got | + | with all the different teams and learn from each other. We got many impressions about other projects and we |
− | hope we could help with our collaborations. | + | hope that we could help with our collaborations. We would like to thank every team that collaborated with us! |
</p> | </p> | ||
</section> | </section> | ||
<hr> | <hr> | ||
<section class="section grid"> | <section class="section grid"> | ||
− | <div class="sub" onclick="popup(' | + | <div class="sub" |
+ | onclick="popup('colony_pictures')"> | ||
<div class="sub-header"> | <div class="sub-header"> | ||
<h1> | <h1> | ||
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</div> | </div> | ||
<div class="sub-content"> | <div class="sub-content"> | ||
− | + | <img src="https://static.igem.org/mediawiki/2019/f/f6/T--Marburg--ColonyPickingButton.svg"> | |
− | + | ||
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</div> | </div> | ||
</div> | </div> | ||
− | <div id=" | + | <div id="colony_pictures" |
+ | class="popup"> | ||
<div class="popup-container"> | <div class="popup-container"> | ||
<div class="popup-header"> | <div class="popup-header"> | ||
− | <h1 class="title">Colony | + | <h1 class="title">The Colony Picture Collaboration</h1> |
− | <button type="button" onclick="hide(' | + | <button type="button" |
+ | onclick="hide('colony_pictures')">X</button> | ||
</div> | </div> | ||
− | <div class="popup-content" style="text-align: justify;"> | + | <div class="popup-content" |
− | <section class="section"> | + | style="text-align: justify;"> |
− | + | <section class="section"> | |
− | + | ||
− | + | ||
− | src=https://static.igem.org/mediawiki/2019/5/5e/T--marburg--colony_picking_2.jpeg | + | <div style="display: flex; flex-direction: row; align-items: center;"> |
− | + | <p><b> | |
− | + | In our colony picking collaboration we asked other iGEM teams to supply us with data for our | |
− | + | picking algorithm. | |
− | + | Thanks to this group effort we could build our Colony Picking Unit.</b> | |
− | + | </p> | |
− | + | ||
− | + | <figure style="text-align: center; margin-left: 1em;"> | |
− | + | <img src="https://static.igem.org/mediawiki/2019/f/f6/T--Marburg--ColonyPickingButton.svg" | |
− | + | alt="ColonyPickingButton" | |
− | + | style="min-width: 200px;"> | |
− | + | ||
− | + | </figure> | |
− | + | </div> | |
− | + | ||
− | + | <div style="display: flex; flex-direction: row;"> | |
− | + | <figure style="text-align: center; margin-right: 1em;"> | |
− | + | <img src=https://static.igem.org/mediawiki/2019/5/5e/T--marburg--colony_picking_2.jpeg | |
− | + | alt="Pipetting Robot, colony picking set up" | |
− | + | style="min-width: 350px;"> | |
− | + | <figcaption> | |
− | + | Figure 1: Pipetting Robot (OT-2) colony picking set up. | |
− | + | </figcaption> | |
− | < | + | </figure> |
− | < | + | <div> |
− | src="https://static.igem.org/mediawiki/2019/f/f8/T--marburg--taking_colony_pictures.png" | + | |
− | + | <p> | |
− | + | Many repetitive tasks in the lab cost time such as pipetting or picking colonies from agar plates | |
− | + | and could technically be performed by robots. | |
− | + | However, most commercially available solutions are simply too expensive and therefore unavailable | |
− | + | for many Labs and iGEM Teams. | |
− | < | + | So one of our goals was to build a reasonable colony picking robot to outsource the tiring task of |
− | + | colony picking by hand. | |
− | < | + | </p> |
− | + | <p> | |
− | < | + | We used an Opentrons OT-2, the cheapest available pipetting robot and constructed modules and |
− | src="https://static.igem.org/mediawiki/2019/0/0b/T--marburg--collonycollab_website.png" | + | scripts, |
− | + | that allow other iGEM Teams to turn their own OT-2 into a colony picking robot as well. | |
− | + | See here how to transfer your OT2 into a | |
− | + | <a style="padding: 0;" | |
− | + | href="https://2019.igem.org/Team:Marburg/Colony_Picking"> Colony Picker. | |
− | + | </a> | |
− | + | ||
− | + | </p> | |
− | + | </div> | |
− | teams that sent us | + | </div> |
− | + | ||
− | + | <div style=" margin-top: 1em;"> | |
− | + | <p> | |
+ | We chose to build an Artificial Intelligence (AI) to automatically recognise colonies. While a “hard-coded” | ||
+ | solution would be easier to achieve, they lack accuracy and flexibility, key attributes of Synbio projects. | ||
+ | The drawback for an AI approach is the amount of data it needs for precise and robust training. | ||
+ | With the help of the iGEM-community we thought to overcome this problem. We asked other iGEM Teams | ||
+ | if they could provide us with labeled pictures of their agar plates with <i>E. coli</i> colonies, which served as the | ||
+ | basis for our AI. For that purpose we set up a | ||
+ | <a style="padding: 0;" | ||
+ | href="https://www.vinca.de/igem/"> Website. | ||
+ | </a> | ||
+ | </p> | ||
+ | </div> | ||
+ | |||
+ | <div style="display: flex; flex-direction: row; margin-top: 1em;"> | ||
+ | <p> | ||
+ | To participate all that was needed were agar plates with <i>E. coli</i> colonies, a screen and a camera. Other | ||
+ | criteria were evenly distributed, distinguishable colonies. The first step was taking the picture. For this | ||
+ | purpose, the screen had to be set to the highest level of brightness and display a white image. When | ||
+ | you take a picture of a common screen, a pattern on the picture could appear, the so called Moiré pattern. | ||
+ | To avoid this, we supplied each team with diffusive sheets, which they could use to cover their screen. | ||
+ | A detailed summary is found <a style="padding: 0;" | ||
+ | href="https://static.igem.org/mediawiki/2019/8/83/T--Marburg--Collaboration.pdf"> here. | ||
+ | </a> | ||
+ | |||
+ | </p> | ||
+ | <figure style="text-align: center; margin-left: 1em;"> | ||
+ | <img src="https://static.igem.org/mediawiki/2019/f/f8/T--marburg--taking_colony_pictures.png" | ||
+ | alt="Pattern for taking the colony picture" | ||
+ | style="min-width: 500px;"> | ||
+ | <figcaption> | ||
+ | Figure 2: Pattern for taking the colony picture. | ||
+ | </figcaption> | ||
+ | </figure> | ||
+ | </div> | ||
+ | |||
+ | <div style="display: flex; flex-direction: row; margin-top: 1em;"> | ||
+ | <figure style="text-align: center; margin-right: 1em;"> | ||
+ | <img src="https://static.igem.org/mediawiki/2019/0/0b/T--marburg--collonycollab_website.png" | ||
+ | alt="colony picking website" | ||
+ | style="min-width: 550px;"> | ||
+ | <figcaption> | ||
+ | Figure 3: Website for colony picture labeling. | ||
+ | </figcaption> | ||
+ | </figure> | ||
+ | <p> | ||
+ | Following this, the images taken had to be labeled using a | ||
+ | <a style="padding: 0;"href="https://www.vinca.de/igem/label/"> web-based | ||
+ | </a> | ||
+ | tool. Afterwards, the single colonies were marked with a rectangle. | ||
+ | In total 250 colony pictures were added to our data set, vastly increasing the performance of our AI | ||
+ | in detecting colonies. As an incentive to participate in this collaboration we gave away prizes to | ||
+ | the teams that sent us their pictures. | ||
+ | </p> | ||
+ | </div> | ||
+ | <div class="wrap-collabsible" | ||
+ | style="margin-top: 25px;"> | ||
+ | <input id="Pizzes" | ||
+ | class="toggle" | ||
+ | type="checkbox"> | ||
+ | <label for="Pizzes" | ||
+ | class="lbl-toggle"> | ||
+ | Participants and Prizes | ||
+ | <hr style="width: unset;"> | ||
+ | </label> | ||
+ | <div class="collapsible-content"> | ||
+ | <div class="content-inner" | ||
+ | style="text-align: left; text-align-last: left;"> | ||
+ | <p style="margin-bottom: 25px;"> | ||
+ | As an incentive to participate in this Collaboration we gave away prizes. The five best teams | ||
+ | that sent us pictures got an <i>E. coli</i> plushie and the three teams that sent us the most | ||
+ | pictures got a bigger surprise. Thanks to Doulix, we provided these three teams with | ||
+ | an engraved trophy out of glass and vouchers. | ||
+ | |||
+ | </p> | ||
+ | <p> | ||
+ | Team <a style="padding: 0;" | ||
+ | href="https://2019.igem.org/Team:BOKU-Vienna"> BOKU-Vienna </a> was able to send us one | ||
+ | picture, team <a style="padding: 0;" | ||
+ | href="https://2019.igem.org/Team:GO_Paris-Saclay"> GO Paris-Saclay</a> provided three pictures | ||
+ | and team <a style="padding: 0;" | ||
+ | href="https://2019.igem.org/Team:TUDelft"> TUDelft</a> could help us with 14 colony | ||
+ | pictures. Team <a style="padding: 0;" | ||
+ | href="https://2019.igem.org/Team:TU_Darmstadt"> TU Darmstadt </a> could send us a number of | ||
+ | 23 pictures, but were beaten by team <a style="padding: 0;" | ||
+ | href="https://2019.igem.org/Team:IISER_Bhopal"> IISER Bhopal </a> which send us an | ||
+ | impressive amount of 99 pictures. | ||
+ | </p> | ||
+ | <figure style="text-align: center; margin-right: 1em;"> | ||
+ | <img src=https://static.igem.org/mediawiki/2019/0/04/T--Marburg--prizes_colony-picture.jpeg | ||
+ | alt="Prizes colony picture"> | ||
+ | <figcaption> | ||
+ | Figure 1: Giving out the prizes for the best participants of the Colony-Picture Project at | ||
+ | the giant Jamboree. | ||
+ | </figcaption> | ||
+ | </figure> | ||
+ | <p> | ||
+ | We thank all the teams for sending us labeled colony pictures. Without your help we would not | ||
+ | have been able to train our AI and to transform our Opentrons OT-2 into a Colony Picker. | ||
+ | </p> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
</section> | </section> | ||
</div> | </div> | ||
</div> | </div> | ||
</div> | </div> | ||
− | <div class="sub" onclick="popup(' | + | <div class="sub" |
+ | onclick="popup('golden_gate')"> | ||
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− | <h1>T H E | + | <h1>T H E   G O L D E N   G A |
T E</h1> | T E</h1> | ||
<h1>C O L L A B O R A T I O N</h1> | <h1>C O L L A B O R A T I O N</h1> | ||
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− | + | <img src="https://static.igem.org/mediawiki/2019/7/71/T--Marburg--GoldenGateCollabosButtons.svg"> | |
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</div> | </div> | ||
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− | <div id=" | + | <div id="golden_gate" |
+ | class="popup"> | ||
<div class="popup-container"> | <div class="popup-container"> | ||
<div class="popup-header"> | <div class="popup-header"> | ||
<h1 class="title">The Golden Gate collaboration</h1> | <h1 class="title">The Golden Gate collaboration</h1> | ||
− | <button type="button" onclick="hide(' | + | <button type="button" |
+ | onclick="hide('golden_gate')">X</button> | ||
</div> | </div> | ||
− | + | <div class="popup-content" | |
− | < | + | style="text-align: justify;"> |
− | + | <section class="section"> | |
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− | + | <div style="display: flex; flex-direction: row; align-items: center;"> | |
− | + | <p><b> | |
− | + | Our team made itself the goal to introduce other iGEM Teams to Golden Gate. | |
− | + | Therefore we hosted a webinar, build up a communication platform | |
− | + | and made an Interlab study.</b> | |
− | + | ||
− | + | ||
</p> | </p> | ||
− | </ | + | <figure style="text-align: center; margin-left: 1em;"> |
+ | <img src="https://static.igem.org/mediawiki/2019/7/71/T--Marburg--GoldenGateCollabosButtons.svg" | ||
+ | alt="GoldenGateCollabosButtons" | ||
+ | style="min-width: 200px;"> | ||
+ | </figure> | ||
</div> | </div> | ||
− | <br> | + | |
− | <div class="wrap-collabsible"> | + | <p> |
− | <input id=" | + | Earlier this year the iGEM headquarters announced changes in their part submission system, one of |
− | <label for=" | + | which made us extremely happy: BioBricks are now also accepted in Type IIS standard! In our opinion, |
− | + | this is a huge step, as cloning techniques based on Type IIS restriction enzymes are becoming more | |
+ | and more popular in the scientific community due to their numerous advantages over classical cloning | ||
+ | methods.<br> | ||
+ | We noticed that more teams converted to MoClo systems in their projects this year but we still think | ||
+ | there is a lot that needs to be done for the iGEM community to really embrace this change. In order | ||
+ | to help teams with the transition into this new system, we had an idea: | ||
+ | What if we could help other teams understand what Type IIS enzymes really are and how they work? If | ||
+ | we could share our passion for Modular Cloning and the value of the PhytoBrick standard? If we could | ||
+ | introduce them to our Marburg Collection and help removing some of the burdens that come with a | ||
+ | switch to a new cloning system?<br> | ||
+ | Therefore we hosted a webinar about Golden Gate, build up a communication platform to give iGEM | ||
+ | Teams a chance to troubleshoot their methods and made an Interlab study. Here we evaluated the | ||
+ | consistency of Golden Gate Assembly protocols across iGEM teams as well as to give teams the | ||
+ | opportunity to gain experience with the assembly method. | ||
+ | </p> | ||
+ | </section> | ||
+ | <section class="section"> | ||
+ | <div class="wrap-collabsible" | ||
+ | style="margin-bottom: 25px;"> | ||
+ | <input id="collapsible2_1" | ||
+ | class="toggle" | ||
+ | type="checkbox"> | ||
+ | <label for="collapsible2_1" | ||
+ | class="lbl-toggle"> | ||
+ | Webinar - Educating about Golden Gate cloning | ||
+ | |||
+ | |||
+ | <hr style="width: unset;"> | ||
</label> | </label> | ||
<div class="collapsible-content"> | <div class="collapsible-content"> | ||
− | <div class="content-inner" style="text-align: | + | <div class="content-inner" |
− | <div> | + | style="text-align: justify; text-align-last: left;"> |
+ | <div style="display: flex; flex-direction: row;"> | ||
+ | |||
+ | <figure style="text-align: center; margin-right: 25px; min-width:200px"> | ||
+ | <img src="https://static.igem.org/mediawiki/2019/3/37/T--Marburg--WebinarButton.svg" | ||
+ | alt="Webinarbutton"> | ||
+ | </figure> | ||
+ | |||
<p> | <p> | ||
− | + | We wanted to reach as many teams as possible with our webinar on Golden Gate cloning, | |
− | + | Type IIS enzymes, their benefits and how to use all of this in their own projects. We | |
− | + | started planning everything that we thought should be in the webinar, from the | |
− | + | biological function of the Type IIS enzymes and how they differ from Type II enzymes, | |
− | + | over its applications in Synthetic Biology like modular cloning to the in-depth | |
− | + | strategies when designing your own parts and actually building them in the lab.<br> <br> | |
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− | designing | + | |
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− | + | </div> | |
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<p> | <p> | ||
− | + | During this planning step we realized, that some of the teams that are already working | |
+ | with Golden Gate might not benefit as much from an entry level webinar. Consequently, we | ||
+ | thought about integrating more complex design questions that we could share with the | ||
+ | community. We finished our webinars with a practical part, showing the part-design in | ||
+ | Geneious Prime – the sequence analysis software we used throughout the year. Firstly, we | ||
+ | explained the different levels that can be built with Golden Gate and then introduced | ||
+ | some more complex designs that are also possible with Golden Gate cloning, such as the | ||
+ | construction of whole operons or space holder parts. Through this we hoped to also reach | ||
+ | teams that use Golden Gate themselves, showing them the possibilities this cloning | ||
+ | system offers.<br> | ||
+ | |||
+ | </p> | ||
+ | <div style="display: flex; flex-direction: row; margin-top: 25px"> | ||
+ | |||
<p> | <p> | ||
− | + | To reach as many teams on the globe as possible we set up webinars on three different | |
− | + | times to cover as many time zones within a comfortable time frame – hoping that this | |
− | + | will allow more iGEM teams to participate. | |
− | + | The one and a half hour webinars were held on July 30th 10am CET, July 31st 6pm CET and | |
− | + | the last time on 8th August 12pm CET by our instructor René Inckemann with the support | |
− | + | of three of our team members. | |
− | + | We were happy that in total 18 teams participated, as well as some students from the | |
− | + | universities of Marburg and Gießen and even a few fellow scientists from the | |
− | + | Max-Planck-Institute for terrestrial microbiology. In total we counted around 70 | |
− | + | participants. We were also really happy to see Sota Hirano from Doulix on our webinar, | |
− | + | who gave us some awesome feedback, stating that he did learn new things about Golden | |
− | + | Gate which he was not aware of before! | |
− | + | </p> | |
− | + | <figure style="text-align: center; margin-left: 25px; min-width:400px"> | |
− | + | <img src="https://static.igem.org/mediawiki/2019/f/f1/T--marburg--Doulix_webinar.jpeg" | |
− | + | alt="Bilder Webinar" | |
− | + | style="max-height: 600px;"> | |
− | + | <figcaption> | |
− | + | Figure 3: Participants watching our Golden Gate webinar all over the world. | |
− | + | </figcaption> | |
− | + | </figure> | |
− | + | </div> | |
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− | + | <div style="display: flex; flex-direction: row; margin-top: 25px"> | |
− | + | <div> | |
− | + | <p> | |
− | + | If you would like to learn about Golden Gate assembly please feel free to watch our | |
− | + | webinar through this Link: | |
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</p> | </p> | ||
+ | <div class="content"> | ||
+ | <ul> | ||
+ | <li><a href="https://www.youtube.com/watch?v=-UUUG2iL7og">Webinar 30.07.2019</a></li> | ||
+ | <li><a href="https://www.youtube.com/watch?v=Aq-6MlaVWww">Webinar 31.07.2019</a></li> | ||
+ | <li><a href="https://www.youtube.com/watch?v=Aq-6MlaVWww">Webinar 09.08.2019</a></li> | ||
+ | </ul> | ||
+ | </div> | ||
+ | <p> | ||
+ | We thank all the following teams for participating in this awesome event: | ||
+ | </p> | ||
+ | <div class="content"> | ||
+ | <ul> | ||
+ | <li><a href="https://2019.igem.org/Team:Aachen/collaborations">Team Aachen</a></li> | ||
+ | <li><a href="https://2019.igem.org/Team:Athens/Collaborations">Team Athens</a></li> | ||
+ | <li><a href="https://2019.igem.org/Team:Amazonas-Brazil/Collaborations">Team | ||
+ | Amazonas-Brazil</a></li> | ||
+ | <li><a href="https://2019.igem.org/Team:CMUQ/Collaborations">Team CMU-Q</a></li> | ||
+ | <li><a href="https://2019.igem.org/Team:ITESO_Guadalajara/Collaborations">Team ITESO</a> | ||
+ | </li> | ||
+ | <li><a href="https://2019.igem.org/Team:SoundBio//Collaborations">Team Soundbio</a></li> | ||
+ | <li><a href="https://2019.igem.org/Team:UFPR-Curitiba-Brazil/Collaborations">Team | ||
+ | UFPR-Curitiba Brazil</a></li> | ||
+ | <li><a href="https://2019.igem.org/Team:TUDelft/Collaborations">Team Delft</a></li> | ||
+ | <li><a href="https://2019.igem.org/Team:BOKU-Vienna/Collaborations">Team BOKU-Viena</a> | ||
+ | </li> | ||
+ | <li><a href="https://2019.igem.org/Team:Duesseldorf/Collaborations">Team Düsseldorf</a> | ||
+ | </li> | ||
+ | <li><a href="https://2019.igem.org/Team:IISc-Bangalore/Collaborations">Team | ||
+ | IlSc-BAngalore</a> | ||
+ | </li> | ||
+ | <li><a href="https://2019.igem.org/Team:TU_Kaiserslautern/Collaborations">Team | ||
+ | Kaiserslautern</a></li> | ||
+ | <li><a href="https://2019.igem.org/Team:Leiden/Collaborations">Team Leiden</a></li> | ||
+ | <li><a href="https://2019.igem.org/Team:UM_Macau/Collaborations">Team UM Macau</a></li> | ||
+ | <li><a href="https://2019.igem.org/Team:PuiChing_Macau/Collaborations">Team Pui Ching | ||
+ | Macau</a></li> | ||
+ | <li><a href="https://2019.igem.org/Team:Tartu_TUIT/Collaboration">Team FAU-Tartz-TUIT</a> | ||
+ | </li> | ||
+ | <li><a href="(https://2019.igem.org/Team:FAU_Erlangen/Collaborations">Team | ||
+ | FAU-Erlangen</a> | ||
+ | </li> | ||
+ | <li><a href="https://2019.igem.org/Team:Queens_Canada/Collaborations">Team | ||
+ | Quens-Canada</a> | ||
+ | </li> | ||
+ | </ul> | ||
+ | </div> | ||
+ | |||
</div> | </div> | ||
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</div> | </div> | ||
</div> | </div> | ||
</div> | </div> | ||
− | <div class=" | + | <div class="wrap-collabsible" |
− | < | + | style="margin-bottom: 25px;"> |
− | + | <input id="collapsible2_2" | |
− | + | class="toggle" | |
− | + | type="checkbox"> | |
− | + | <label for="collapsible2_2" | |
− | + | class="lbl-toggle"> | |
− | + | Slack Channel and gDrive - Global networking in the Golden Gate community | |
− | + | <hr style="width: unset;"> | |
− | + | </label> | |
− | + | <div class="collapsible-content"> | |
− | + | <div class="content-inner" | |
− | + | style="text-align: left; text-align-last: left;"> | |
− | < | + | |
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− | <div class=" | + | |
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− | + | ||
− | + | ||
− | + | <figure style="text-align: center; margin-right: 25px; min-width:200px"> | |
− | + | <img src="https://static.igem.org/mediawiki/2019/3/30/T--Marburg--GoldenGateCommunicationButton.svg" | |
− | + | alt="GoldenGateCommunicationButton"> | |
− | + | </figure> | |
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− | + | ||
− | + | ||
− | + | <p style="margin-bottom: 25px;"> | |
− | + | Although we were quite pleased with the result of our webinar, we thought about how to | |
− | + | further connect this Golden Gate community, so we created a Slack workspace for everyone | |
− | + | that is working with this method, enabling us all to exchange protocols and ideas or | |
− | + | troubleshoot issues that might arise. So far there are close to 50 members, but we hope | |
− | + | to find more enthusiasts willing to share their knowledge!<br> | |
− | + | </p> | |
− | + | </div> | |
− | + | </div> | |
− | + | ||
− | + | <p style="margin-bottom: 25px;"> | |
− | + | All files used for the webinar are openly available. You can find our protocols, | |
− | + | handouts, slides as well as the files we used for the practical part in Geneious to | |
− | + | construct parts in-silico in our shared Google Drive. Through this we hope to make it | |
− | + | easier for other teams willing to start with Golden Gate cloning, as they can start | |
− | + | designing their first own in-silico parts with our guidance. | |
− | + | ||
− | + | ||
</p> | </p> | ||
− | < | + | <p style="margin-bottom: 25px;"> |
− | + | The main documents of the Golgen Gate Webinar<br> | |
− | < | + | → Download: <a |
− | + | href="https://static.igem.org/mediawiki/2019/0/0a/T--marburg--GG_webinar_slides.pdf">Golden | |
− | + | Gate webinar slide 2019</a><br> | |
− | + | → Download: <a | |
− | + | href="https://static.igem.org/mediawiki/2019/0/09/T--CMUQ--goldenGateWebinarHandout.pdf">Handout | |
− | + | Golden Gate webinar</a><br> | |
− | + | → Download: <a | |
− | + | href="https://static.igem.org/mediawiki/2019/c/c2/T--marburg--GG_protocoll_white.pdf">Golden | |
− | + | Gate protocoll</a> | |
− | + | ||
− | + | ||
− | + | ||
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− | < | + | |
− | + | ||
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− | < | + | |
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− | + | ||
− | </ | + | |
− | + | ||
</p> | </p> | ||
− | |||
<p> | <p> | ||
− | + | To try out the described in-silico cloning you can download the needed sequences as | |
− | + | geneious file here.<br> | |
− | + | → Download: <a href="https://2019.igem.org/File:T--marburg--Sequences_Webinar.zip">Sequences | |
− | + | Webinar </a> | |
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</p> | </p> | ||
</div> | </div> | ||
− | |||
− | |||
</div> | </div> | ||
</div> | </div> | ||
− | <div class=" | + | <div class="wrap-collabsible" |
− | < | + | style="margin-bottom: 25px;"> |
− | + | <input id="collapsible2_3" | |
− | + | class="toggle" | |
− | + | type="checkbox"> | |
− | + | <label for="collapsible2_3" | |
− | + | class="lbl-toggle"> | |
− | + | Interlab - Accelerate Golden Gate Assembly | |
− | + | <hr style="width: unset;"> | |
− | + | </label> | |
− | + | <div class="collapsible-content"> | |
− | < | + | <div class="content-inner" |
− | + | style="text-align: left;"> | |
− | + | <div style="display: flex; flex-direction: row;"> | |
− | + | <p> | |
− | + | ||
− | <div class=" | + | |
− | <div | + | |
− | + | ||
− | + | As we are constantly working on improving the efficiency of Golden Gate. We clearly | |
− | + | saw the main bottleneck of the reaction was the long assembly time in the cycler. | |
− | + | In the webinar and in our Golden Gate community we provided protocols with three different | |
− | + | cycler settings for different applications. | |
− | + | Although these three protocols were tested before, they were never compared to each other in | |
− | + | their efficiency.<br> | |
− | + | We conducted our own Interlab study in collaboration with the iGEM Teams from | |
− | + | <a style="padding: 0;" | |
− | + | href="https://2019.igem.org/Team:TUDelft"> TUDelft </a>, | |
− | + | <a style="padding: 0;" | |
− | + | href="https://2019.igem.org/Team:Humboldt_Berlin"> HUmboldt Berlin </a> | |
− | + | and | |
− | + | <a style="padding: 0;" | |
− | + | href="https://2019.igem.org/Team:TU_Kaiserslautern"> TU Kaiserslautern | |
− | + | </a> | |
− | + | In order to find the best protocol, we supplied the teams with specific Level 0 | |
− | + | parts,that should be assembled to a Level 1 vector using the three different cycler | |
− | + | conditions. The provided parts are shown in the table | |
− | + | </p> | |
− | < | + | <div style="display: flex; flex-direction: column; margin-left: 25px;"> |
− | + | <figure style="float:left;"> | |
− | < | + | <img src="https://static.igem.org/mediawiki/2019/c/c9/T--Marburg--InterlabButton.svg" |
− | + | alt="Interlab button" | |
− | < | + | style="max-height: 200px;"> |
− | src="https://static.igem.org/mediawiki/2019/ | + | </figure> |
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | |||
− | + | <table style="min-width: 400px;"> | |
− | + | <tbody> | |
− | + | <tr> | |
− | + | <td style="text-align: center">5' Connector </td> | |
− | + | <td style="text-align: center"> <a href="link 1"> BBa_K2560075 </a>/ | |
− | + | </tr> | |
− | + | <tr> | |
− | + | <td style="text-align: center">Promoter </td> | |
− | + | <td style="text-align: center"> <a href="link 2"> J23119 BBa_K2560031 </a> | |
− | + | </tr> | |
− | + | <tr> | |
− | < | + | <td style="text-align: center">RBS </td> |
− | + | <td style="text-align: center"> <a href="link 2"> B0034 BBa_K2560008 </a> | |
− | alt=" | + | </tr> |
+ | <tr> | ||
+ | <td style="text-align: center">CDS </td> | ||
+ | <td style="text-align: center"> <a href="link 3"> sfGFP BBa_K2560042 </a> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td style="text-align: center">Terminator </td> | ||
+ | <td style="text-align: center"> <a href="link 4"> B0015 BBa_K2560035 </a> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td style="text-align: center">3' Connector </td> | ||
+ | <td style="text-align: center"> <a href="link 5"> BBa_K2560080 </a> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td style="text-align: center">ORI: </td> | ||
+ | <td style="text-align: center"> <a href="link 6"> ColE1 BBa_K2560036 </a> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td style="text-align: center">Antibiotic-Resistance-Cassette</td> | ||
+ | <td style="text-align: center"> <a href="link 7"> Kan-RFP BBa_K2560056 </a> | ||
+ | </tr> | ||
+ | </tbody> | ||
+ | </table> | ||
+ | </div> | ||
+ | </div> | ||
+ | <div style="display: flex; flex-direction: row;"> | ||
+ | <figure style="text-align: center; margin-right: 25px"> | ||
+ | <img src="https://static.igem.org/mediawiki/2019/9/91/T--marburg--interlab_package.jpeg" | ||
+ | alt="Content of the Interlab package"> | ||
<figcaption> | <figcaption> | ||
− | Fig | + | Fig 1: Content of the Interlab package. |
− | + | ||
</figcaption> | </figcaption> | ||
</figure> | </figure> | ||
+ | <p> | ||
+ | The Golden Gate reactions should be pipetted together using the given parts and then | ||
+ | be transformed in <i>E. coli</i> and be plated with similar amounts on agar plates. A colony with a | ||
+ | correct assembled | ||
+ | vector would have a functioning transcription unit which expresses GFP, which can be clearly | ||
+ | distinguished from RFP expressing colonies which only carries the antibiotic cassette and | ||
+ | false non fluorescent | ||
+ | colonies.<br> | ||
+ | The ratio of green to non-green colonies was used to evaluate which of the three protocols | ||
+ | yielded in the | ||
+ | highest transformation efficiency and the highest rate of positive clones. | ||
+ | </p> | ||
+ | </div> | ||
+ | <p> | ||
+ | The handout with the used protocols can be downloaded <a | ||
+ | href=https://static.igem.org/mediawiki/2019/a/a0/T--marburg--Interlab_protocoll.pdf>here | ||
+ | </a>. | ||
+ | </p> | ||
+ | <p> | ||
+ | <br> | ||
+ | <b>Results</b> | ||
+ | <br> | ||
− | + | <div style="display: flex; flex-direction: row;"> | |
− | + | <p> First we wanted to know which protocol worked best for every team. For this we analysed the data of the correct assembled green colonies for every protocol. The data of the different teams are given in Table 1 and are shown in as chart in Figure 2. The total colony count is hard to compare directly, cause the numbers differ very much between the teams. This could come through the different transformation efficiencies for example because of the variating competent cells the teams used. </p> | |
− | + | ||
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− | + | ||
− | <img style="height: | + | |
− | + | <figure style="text-align: center; margin-left: 25px; min-width:500px"> | |
− | + | <figcaption style="max-width: 500px; text-align: center"> | |
− | + | Table 1 - Data for the gained green colonies gained by using the three different protocols, from each participating team. | |
− | + | </figcaption> | |
− | + | <img | |
− | + | src="https://static.igem.org/mediawiki/2019/4/4a/T--Marburg--Tabelle_Interlab.png" | |
− | + | alt="Table 1 - Data for the gained green colonies gained by using the three different protocols, from each participating team"> | |
− | + | </figure> | |
− | + | <br> | |
+ | </div> | ||
+ | |||
+ | <figure style="text-align:center"> | ||
+ | <img style="height: 400px; width: 500px;" | ||
+ | src="https://static.igem.org/mediawiki/2019/3/3a/T--Marburg--Interlab_1.png" | ||
+ | alt="Amount of gained green colonies gained by using the three different protocols, from each participating team."> | ||
+ | <figcaption style="max-width: 2400px; text-align: center"> | ||
+ | Fig.2 Amount of gained green colonies gained by using the three different protocols, from each participating team | ||
+ | </figcaption> | ||
+ | </figure> | ||
+ | |||
+ | <p> | ||
+ | <br> | ||
+ | The results in Figure 2 show, that the protocols work differently for each team. In general the most colonies were | ||
+ | received with the long Troubleshoot protocol, but also the Rapid and Improved protocols show high numbers of correct colonies. | ||
+ | <br> | ||
+ | But which protocol is the most efficient one? | ||
+ | To compare the results of the teams the ratio of the gained correct colonies between the | ||
+ | protocols were averaged. The data are shown in Figure 3 and show, that the long Troubleshoot | ||
+ | assembly works the best. The fastest assembly (Rapid) works the second best. This is very | ||
+ | interesting, because this assembly takes significant less time.<br> | ||
+ | <br> | ||
+ | We would propose to take the Troubleshoot protocol for complicated assemblies and the rapid | ||
+ | protocol for easy assemblies like lvl 0.<br> | ||
+ | </p> | ||
+ | <figure style="text-align:center"> | ||
+ | <img style="height: 400px; width: 500px;" | ||
+ | src="https://static.igem.org/mediawiki/2019/a/a1/T--Marburg--Interlab_3.png" | ||
+ | alt="Fig.3 Average efficiency of the three protocols"> | ||
+ | <figcaption style="max-width: 2400px; text-align: center"> | ||
+ | Fig.3 Average efficiency of the three protocols. | ||
+ | </figcaption> | ||
+ | </figure> | ||
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</div> | </div> | ||
+ | </section> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | <div class="sub" | ||
+ | onclick="popup('berlin')"> | ||
+ | <div class="sub-header"> | ||
+ | <h1>B E R L I N</h1> | ||
+ | <hr> | ||
+ | </div> | ||
+ | <div class="sub-content"> | ||
+ | <img src="https://static.igem.org/mediawiki/2019/9/9b/T--marburg--berlin_logo_richtig.png"> | ||
+ | </div> | ||
+ | |||
+ | </div> | ||
+ | <div id="berlin" | ||
+ | class="popup"> | ||
+ | <div class="popup-container"> | ||
+ | <div class="popup-header"> | ||
+ | <h1 class="title">Berlin Collaboration</h1> | ||
+ | <button type="button" | ||
+ | onclick="hide('berlin')">X</button> | ||
+ | </div> | ||
+ | <div class="popup-content" | ||
+ | style="text-align: justify;"> | ||
+ | <div style="display: flex; flex-direction: row;"> | ||
+ | <p> | ||
+ | iGEM Berlin engineered the freshwater algae <i>Chlamydomonas reinhardtii</i> to degrade | ||
+ | Polyethylene | ||
+ | terephthalate | ||
+ | (PET). For this goal they also built a bioreactor for their chassis. We sent | ||
+ | them our UTEX | ||
+ | strain to test if | ||
+ | their photobioreactor is capable of cultivating other phototrophic organisms as | ||
+ | well. They gained | ||
+ | additional | ||
+ | experiences on growth conditions for phototrophic organisms and we could see our | ||
+ | strains grow in | ||
+ | a bioreactor environment.<br> | ||
+ | Click here to learn more about there project <a | ||
+ | href="https://2019.igem.org/Team:Humboldt_Berlin">Chlamylicious</a> | ||
+ | </p> | ||
+ | <figure style="float:right; margin-left: 25px;"> | ||
+ | <img src="https://static.igem.org/mediawiki/2019/9/9b/T--marburg--berlin_logo_richtig.png" | ||
+ | alt="Berlin logo" | ||
+ | style="min-width: 500px;"> | ||
+ | </figure> | ||
</div> | </div> | ||
− | + | </div> | |
+ | </div> | ||
+ | </div> | ||
+ | <div class="sub" | ||
+ | onclick="popup('delft')"> | ||
+ | <div class="sub-header"> | ||
+ | <h1>D E L F T</h1> | ||
+ | <hr> | ||
+ | </div> | ||
+ | <div class="sub-content"> | ||
+ | <img src="https://static.igem.org/mediawiki/2019/d/d4/T--marburg--DELF_logo_richtig.svg"> | ||
+ | </div> | ||
+ | </div> | ||
+ | <div id="delft" | ||
+ | class="popup"> | ||
+ | <div class="popup-container"> | ||
+ | <div class="popup-header"> | ||
+ | <h1 class="title">Delft Collaboration</h1> | ||
+ | <button type="button" | ||
+ | onclick="hide('delft')">X</button> | ||
+ | </div> | ||
+ | <div class="popup-content" | ||
+ | style="text-align: justify;"> | ||
+ | <div style="display: flex; flex-direction: row; margin-top: 25px;"> | ||
+ | <figure style="margin-right: 25px;"> | ||
+ | <img src="https://static.igem.org/mediawiki/2019/5/5e/T--marburg--Skype_DELF.jpeg" | ||
+ | alt=" DELFT Bild" | ||
+ | style="max-width: 300px;"> | ||
+ | <figcaption> Fig. 1: Skype talk with TU Delft</figcaption> | ||
+ | </figure> | ||
+ | <div style="display: flex; flex-direction: column;"> | ||
+ | <div> | ||
+ | <h2 class="subtitle">Delft Collaborations</h2> | ||
+ | <p> | ||
+ | iGEM Delft created an all species encompassing toolbox, based on orthogonal gene | ||
+ | expression and | ||
+ | host | ||
+ | independent plasmid replication. We exchanged multiple times over Molecular | ||
+ | Cloning and toolbox | ||
+ | designs. | ||
+ | Additionally, we sent them parts in the PhytoBricks standard.<br> | ||
+ | Click here to learn more about there project <a href="https://2019.igem.org/Team:TUDelft"> | ||
+ | project SCI-PHI 29 </a> | ||
+ | </p> | ||
+ | </div> | ||
+ | <figure> | ||
+ | <img src="https://static.igem.org/mediawiki/2019/d/d4/T--marburg--DELF_logo_richtig.svg" | ||
+ | alt="DELF logo" | ||
+ | style="max-height: 200px; margin-top: 25px;"> | ||
+ | </figure> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | <div class="sub" | ||
+ | onclick="popup('uc_davis')"> | ||
+ | <div class="sub-header"> | ||
+ | <h1> | ||
+ | U C   D A V I S | ||
+ | </h1> | ||
+ | <hr> | ||
+ | </div> | ||
+ | <div class="sub-content" | ||
+ | style="justify-content: center;"> | ||
+ | <img style="max-height: 100%;" | ||
+ | src="https://static.igem.org/mediawiki/2019/d/d7/T--UC_Davis--Teamlogolight.png"> | ||
+ | </div> | ||
+ | </div> | ||
+ | <div id="uc_davis" | ||
+ | class="popup"> | ||
+ | <div class="popup-container"> | ||
+ | <div class="popup-header"> | ||
+ | <h1 class="title">UC Davis collaboration</h1> | ||
+ | <button type="button" | ||
+ | onclick="hide('uc_davis')">X</button> | ||
+ | </div> | ||
+ | <div class="popup-content"> | ||
+ | <div style="display: flex; flex-direction: row;"> | ||
+ | |||
+ | <div> | ||
+ | <p style="margin-bottom: 25px;"> | ||
+ | UC Davis iGEM team is one of the winners of the Opentrons competition in 2019. | ||
+ | iGEM Marburg | ||
+ | won one OT-2 | ||
+ | during last | ||
+ | year’s edition of the competition and got contacted for a collaboration by iGEM | ||
+ | UC Davis | ||
+ | along with other | ||
+ | winners from | ||
+ | 2018 and 2019. | ||
+ | The goal for this automation InterLab collaboration was to collect data to | ||
+ | determine the | ||
+ | accuracy and | ||
+ | reproducibility of | ||
+ | the OT-2 compared to humans. To do this, UC Davis proposed to perform the | ||
+ | standard iGEM | ||
+ | protocol on serial | ||
+ | dilutions | ||
+ | with silica beads and fluorescein, each manually and with an OT-2. In total iGEM | ||
+ | Marburg did | ||
+ | four dilutions; | ||
+ | twice with | ||
+ | an OT-2 and twice manually. The data was then sent to UC Davis via the | ||
+ | spreadsheet designed | ||
+ | by them. | ||
+ | Finally, iGEM | ||
+ | Marburg aided by creating a Slack channel for all iGEM teams with an OT-2 to | ||
+ | facilitate | ||
+ | better communication | ||
+ | and | ||
+ | exchange in the future. | ||
+ | </p> | ||
+ | <center><figure Style="text-align:center"> | ||
+ | <!-- add this for centered pics--> | ||
+ | <img src="https://static.igem.org/mediawiki/2019/4/49/T--marburg--UCD_OT2.jpeg" width="450" height="300" | ||
+ | alt="UCD OT2"> | ||
+ | <figcaption> | ||
+ | Figure 1: OT-2 setup for the UC Davis collaboration | ||
+ | </figcaption> | ||
+ | </figure></center> | ||
+ | <p> | ||
+ | In total, seven teams participated and sent their data to UC Davis which are | ||
+ | shown in the | ||
+ | figure below. | ||
+ | According to UC | ||
+ | Davis, the fluorescence data was standardized using the iGEM MEFL conversion to | ||
+ | standardize | ||
+ | the plate reader | ||
+ | measurements. Subsequently, the data was fitted using a linear regression model, | ||
+ | whose | ||
+ | quality is quantified | ||
+ | using the | ||
+ | r2 metric (it ranges from 0 to 1 with higher value indicates a better fit). | ||
+ | <br><br> | ||
+ | For the fluorescence the r2 value from the OT2 data beat human r2 value with | ||
+ | 0.94 and 0.90, | ||
+ | respectively. | ||
+ | Interestingly, for the silica beads, manual measurements yielded a more | ||
+ | satisfactory result | ||
+ | with r2 value of | ||
+ | 0.90 | ||
+ | compared to 0.82 from OT2 measurements. One possible explanation could be the | ||
+ | speed of the | ||
+ | OT-2: it took | ||
+ | twice as long | ||
+ | to complete this task with the OT-2 compared to a manual trial. This could lead | ||
+ | to the | ||
+ | sedimentation of the | ||
+ | silica beads | ||
+ | as the OT-2 went through their runs; therefore worsening its results. Also, as | ||
+ | shown in Fig. | ||
+ | 2b, one team’s | ||
+ | result | ||
+ | deviates a lot from the rest of the teams, maybe if their result is taken out from | ||
+ | the overall | ||
+ | evaluation a more | ||
+ | comparable | ||
+ | result between OT-2 and human could be observed. | ||
+ | </p> | ||
+ | </div> | ||
+ | </div> | ||
+ | <p style="margin-top: 25px;"> | ||
+ | More information about this collaboration such as individual team data can be | ||
+ | found on <a href="https://2019.igem.org/Team:UC_Davis/Collaborations"> UC Davis wiki. | ||
+ | </a> | ||
+ | |||
+ | </p> | ||
+ | <figure style="text-align: center; margin-top: 25px;"> | ||
+ | <img style="max-height: 450px" | ||
+ | src="https://static.igem.org/mediawiki/2019/c/cc/T--marburg--UC_Davis_Graphic_1.png" | ||
+ | alt=" UCD Graphic 1"> | ||
+ | <img style="max-height: 450px" | ||
+ | src=" https://static.igem.org/mediawiki/2019/a/a7/T--marburg--UC_Davis_Graphic_2.png" | ||
+ | alt=" UC Davis Graphic 2"> | ||
+ | <figcaption> | ||
+ | Figure 2: a) OT-2 vs manual fluorescein measurement experiment, OT-2 | ||
+ | performed better | ||
+ | with r2 value of 0.94 compared to 0.90 of human. b) OT-2 vs manual silica | ||
+ | beads measurement experiment, human performed better | ||
+ | with r2 value of 0.90 compared to 0.82 of OT-2. | ||
+ | </figcaption> | ||
+ | </figure> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | <div class="sub" | ||
+ | onclick="popup('uflorida')"> | ||
+ | <div class="sub-header"> | ||
+ | <h1> | ||
+ | U F L O R I D A | ||
+ | </h1> | ||
+ | <hr> | ||
+ | </div> | ||
+ | <div class="sub-content"> | ||
+ | <img src="https://static.igem.org/mediawiki/2019/f/f7/T--Florida--iGEMWeb1.png"> | ||
+ | </div> | ||
+ | </div> | ||
+ | <div id="uflorida" | ||
+ | class="popup"> | ||
+ | <div class="popup-container"> | ||
+ | <div class="popup-header"> | ||
+ | <h1 class="title">UFlorida</h1> | ||
+ | <button type="button" | ||
+ | onclick="hide('uflorida')">X</button> | ||
+ | </div> | ||
+ | <div class="popup-content" | ||
+ | style="text-align: justify"> | ||
+ | <div style="display: flex; flex-direction: row;"> | ||
+ | <figure style="float:left; margin-right: 25px;"> | ||
+ | <img src="https://static.igem.org/mediawiki/2019/f/f7/T--Florida--iGEMWeb1.png" | ||
+ | alt="Florida"> | ||
+ | </figure> | ||
+ | <p> | ||
+ | This semester we had the pleasure of mentoring UFlorida for their Wiki | ||
+ | design. Our member, René Inckemann, aided UFlorida in their quest to create | ||
+ | a visually accessible Wiki, similar to what we did this past iGEM season. | ||
+ | We helped shape UFlorida’s website into an interactive and appealing | ||
+ | landscape, while maintaining the integrity of their goal and project. We | ||
+ | advised them on their font choices- specifically their use of bold and | ||
+ | italicized words-, along with answering questions about our work with the | ||
+ | visually disabled community and utilizing our “Accessible Wiki Guide” from | ||
+ | the previous year. | ||
+ | <br> | ||
+ | Besides answering questions specifically about their project, we also | ||
+ | offered valuable information about when to start working on the Wiki and | ||
+ | questions about coding. Working with a young team like UFlorida offered us | ||
+ | a great mentorship opportunity. | ||
+ | |||
+ | <a style="padding: 0;" | ||
+ | href="https://2019.igem.org/Team:Florida/Collaborations"> Click here to learn more about there project. | ||
+ | </a> | ||
+ | |||
+ | |||
+ | </p> | ||
+ | </div> | ||
+ | <figure style="align-content:center; margin-top: 25px;"> | ||
+ | <img src="https://static.igem.org/mediawiki/2019/2/29/T--marburg--UFlorida_Rene.jpeg" | ||
+ | alt="Florida"> | ||
+ | <figcaption> | ||
+ | Fig. 1: Skypecall with UFlorida | ||
+ | </figcaption> | ||
+ | </figure> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | <div class="sub" | ||
+ | onclick="popup('interaction')"> | ||
+ | <div class="sub-header"> | ||
+ | <h1>I N T E R A C T I O N S</h1> | ||
+ | <h1>W I T H</h1> | ||
+ | <h1>O T H E R   T E A M S</h1> | ||
+ | <hr> | ||
+ | </div> | ||
+ | <div class="sub-content"> | ||
+ | <img src="https://static.igem.org/mediawiki/2019/f/fe/T--marburg--Postcard.png"> | ||
+ | </div> | ||
+ | </div> | ||
+ | <div id="interaction" | ||
+ | class="popup"> | ||
+ | <div class="popup-container"> | ||
+ | <div class="popup-header"> | ||
+ | <h1 class="title">Interactions with other teams</h1> | ||
+ | <button type="button" | ||
+ | onclick="hide('interaction')">X</button> | ||
+ | </div> | ||
+ | <div class="popup-content" | ||
+ | style="text-align: justify;"> | ||
+ | <h2 class="subtitle">Mike the microbe</h2> | ||
+ | <div style="display: flex; flex-direction: row;"> | ||
+ | <figure Style="text-align: center; margin-right: 25px;"> | ||
+ | <img src="https://static.igem.org/mediawiki/2019/b/b0/T--marburg--Mike_microbe.png" | ||
+ | alt=" Mike the Mircrobe" | ||
+ | style="min-width: 300px;"> | ||
+ | <figcaption> | ||
+ | Figure 1: Mike the mircrobe in our lab. | ||
+ | </figcaption> | ||
+ | </figure> | ||
+ | <p> | ||
+ | We participated in the “Mike the microbe” collaboration of the iGEM Team US AFRL | ||
+ | CarrollHS. | ||
+ | They | ||
+ | sent us | ||
+ | their virtual mascots Mike the Microbe and Chia the Chitinase so we could take pictures | ||
+ | with | ||
+ | them to | ||
+ | connect | ||
+ | all iGEM teams around the world via social media. It brought us great joy to join to this | ||
+ | collaboration and | ||
+ | let the two little buddies visit our daily life in our incubators for cyanobacteria. | ||
+ | </p> | ||
+ | </div> | ||
+ | <h2 class="subtitle" | ||
+ | style="margin-top: 25px;">Postcards Düsseldorf</h2> | ||
+ | <div style="display: flex; flex-direction: row;"> | ||
+ | <p> | ||
+ | iGEM Düsseldorf called for their traditional collaboration project, a long year postcard | ||
+ | campaign with the | ||
+ | purpose to educate and to show the public about the different topics and projects in | ||
+ | Synthetic | ||
+ | Biology and | ||
+ | especially regarding all the iGEM projects. With interesting designs and a short project | ||
+ | abstract iGEM teams | ||
+ | could spread their ideas using the postcards to share their thinking to each other and the | ||
+ | society. Our | ||
+ | title and postcard design was about our photosynthetic project „Syntex“. You can see | ||
+ | <i>Synechococcus elongatus</i> | ||
+ | UTEX 2973 direly needing a refreshing drink, a cute little draft from our team member | ||
+ | Jonas. | ||
+ | </p> | ||
+ | <figure style="text-align:center; margin-left: 25px;"> | ||
+ | <img src="https://static.igem.org/mediawiki/2019/f/fe/T--marburg--Postcard.png" | ||
+ | alt="Postcard challenge" | ||
+ | style="min-width: 300px;"> | ||
+ | <figcaption> | ||
+ | Figure 2: Our Postcard for the Postcard collaboration from Düsseldorf. | ||
+ | </figcaption> | ||
+ | </figure> | ||
+ | </div> | ||
+ | <h2 class="subtitle" | ||
+ | style="margin-top: 25px;">iGEMxSDGs challenge</h2> | ||
+ | <div style="display: flex; flex-direction: row;"> | ||
+ | <figure style="text-align:center; margin-right: 25px;"> | ||
+ | <img src="https://static.igem.org/mediawiki/2019/1/18/T--marburg--SDG_chalenge.png" | ||
+ | alt="SGD Challenge - 7 12 13 15" | ||
+ | style="min-width: 300px;"> | ||
+ | <figcaption> | ||
+ | Figure 3: SDG challenge. | ||
+ | </figcaption> | ||
+ | </figure> | ||
+ | <p> | ||
+ | IGEM Costa Rica, Tuebingen, and TAS Taipei created an iGEMxSDGs challenge as | ||
+ | collaboration. | ||
+ | They | ||
+ | asked us to | ||
+ | choose four of the Sustainable Development Goals (SDGs) proposed by the United Nations | ||
+ | (UN). | ||
+ | We | ||
+ | joined this | ||
+ | collaboration challenge and spread it on our social media account. Our project targets the | ||
+ | goals shown in Figure 3. | ||
+ | </p> | ||
+ | </div> | ||
+ | <h2 class="subtitle" | ||
+ | style="margin-top: 25px;">Who wore it best? #labcoatchallenge</h2> | ||
+ | <div style="display: flex; flex-direction: row;"> | ||
+ | <p> | ||
+ | The Stony Brook iGEM Team challenged us to design an outfit for a runway from a standard | ||
+ | lab | ||
+ | coat. | ||
+ | The | ||
+ | outfit had to be created by tying, not using duct tape, cut or sew, etc. After designing | ||
+ | our | ||
+ | outfits | ||
+ | four of | ||
+ | our members presented their style on our ‘runway’ while paparazzi took their photos. As an | ||
+ | outfit we | ||
+ | chose a | ||
+ | dress highlighting the neckline, a shoulder free dress, a knotted dress and a skirt to | ||
+ | bring | ||
+ | glamour | ||
+ | to the | ||
+ | daily routine in the labs. Especially our guys were rocking their outfits as you can see | ||
+ | in Figure 4. | ||
+ | </p> | ||
+ | <figure style="text-align:center; margin-left: 25px;"> | ||
+ | <img src="https://static.igem.org/mediawiki/2019/7/74/T--marburg--Labcoat_Burgi.jpeg" | ||
+ | alt="Our model Burgis for the labcoatchallenge" | ||
+ | style="min-width: 300px;"> | ||
+ | <figcaption> | ||
+ | Figure 4: Our model Burgis for the labcoatchallenge. | ||
+ | </figcaption> | ||
+ | </figure> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | <div class="sub" | ||
+ | onclick="popup('meetups')"> | ||
+ | <div class="sub-header"> | ||
+ | <h1>M E E T U P S</h1> | ||
+ | <hr> | ||
+ | </div> | ||
+ | <div class="sub-content"> | ||
+ | <img src="https://static.igem.org/mediawiki/2019/9/90/T--Marburg--meetups.jpg"> | ||
+ | </div> | ||
+ | </div> | ||
+ | <div id="meetups" | ||
+ | class="popup"> | ||
+ | <div class="popup-container"> | ||
+ | <div class="popup-header"> | ||
+ | <h1 class="title">Meet ups</h1> | ||
+ | <button type="button" | ||
+ | onclick="hide('meetups')">X</button> | ||
+ | </div> | ||
+ | <div class="popup-content" | ||
+ | style="text-align: justify;"> | ||
+ | <h2 class="subtitle">iGEM Spring Festival in Bonn</h2> | ||
+ | <div style="display: flex; flex-direction: row;"> | ||
+ | <p> | ||
+ | From the 3rd to the 5th Mai we met other iGEMers at the Spring Festival in Bonn. Professors | ||
+ | of the university of Bonn presented their research fields which was met with huge interest. We heard | ||
+ | lectures | ||
+ | about immunology and neurosciences, as well as reviving mammoths. During the lectures, | ||
+ | specific | ||
+ | topics were deepened in small seminar groups, such as the ethical tenability of CRISPR | ||
+ | applied | ||
+ | at humans and other animals. At the poster sessions we got the first time in contact with | ||
+ | other teams’ projects. Afterwards, we had the opportunity to vote for the best project, | ||
+ | which | ||
+ | iGEM Düsseldorf won. Besides that, the meetup was filled with a lot of awarding | ||
+ | categories. | ||
+ | We | ||
+ | competed in the categories for “Best Video”, “Best Poster”, “Best Project” and a single | ||
+ | person | ||
+ | had the chance to win the award for “Best Meme”. The iGEM Team Bonn offered spontaneously | ||
+ | a | ||
+ | little football table tournament, where two of our team members won. In the evening we | ||
+ | learnt a | ||
+ | lot about Bonn as a city from a guide living there for many years. Subsequently, the iGEM Teams | ||
+ | met | ||
+ | in | ||
+ | several groups in restaurants and bars. We merged with iGEM Kaiserslautern and exchanged | ||
+ | our | ||
+ | projects and ideas for the rest of the evening.<br> | ||
+ | On the second day, the iGEM Team Bonn offered a competition out of three rounds, where we | ||
+ | competed against other teams. We were asked iGEM specific questions, had to solve riddles | ||
+ | for an | ||
+ | escape room and finished a lab challenge afterwards. Our team could compete until the end | ||
+ | and | ||
+ | won the prize! Thanks again to Doulix for this awesome prize! <br> | ||
+ | We particularly thank the iGEM Team Bonn for the huge effort and the great meetup! | ||
+ | </p> | ||
+ | <figure style="text-align:center; margin-left: 25px"> | ||
+ | <img style="min-width: 300px;" | ||
+ | src="https://static.igem.org/mediawiki/2019/a/a9/T--marburg--meet_u_bonn.png" | ||
+ | alt="Erlangen meetup"> | ||
+ | <figcaption> | ||
+ | Fig. 1: Two representatives of Doulix celebrating with us. | ||
+ | </figcaption> | ||
+ | </figure> | ||
+ | </div> | ||
+ | <h2 class="subtitle" | ||
+ | style="margin-top: 25px;">Erlangen collaboration meetup</h2> | ||
+ | <div style="display: flex; flex-direction: row;"> | ||
+ | <figure style="text-align:center; margin-right: 25px"> | ||
+ | <img style="min-width: 300px;" | ||
+ | src="https://static.igem.org/mediawiki/2019/0/01/T--marburg--meet_up_erlangen.jpeg" | ||
+ | alt="Erlangen meetup"> | ||
+ | <figcaption> | ||
+ | Fig. 2: Group photo from the Erlangen mett up (Michael Burgis from Team Marburg in the | ||
+ | left | ||
+ | side). | ||
+ | </figcaption> | ||
+ | </figure> | ||
+ | <p> | ||
+ | On the 17th till the 19th of May we attended the collaboration meetup hosted by the iGEM team | ||
+ | Erlangen | ||
+ | 2019. There, we met the iGEM teams of Straßbourg, Tübingen, Vienna and of course Erlangen. | ||
+ | Apart | ||
+ | from listening to the interesting talks from the various professors there, we made a lot | ||
+ | of | ||
+ | new | ||
+ | friends and took the chance to talk about possible collaborations with the other teams and | ||
+ | also | ||
+ | promote our collaborations, such as the <a | ||
+ | href="https://2019.igem.org/Team:Marburg/Collaborations#colony_pictures" | ||
+ | target="_blank">colony picking project</a> and the <a | ||
+ | href="https://2019.igem.org/Team:Marburg/Collaborations#golden_gate" | ||
+ | target="_blank">Golden Gate webinar</a>. We would like to thank iGEM team Erlangen very much for | ||
+ | organizing this | ||
+ | wonderful meetup! | ||
+ | </p> | ||
+ | </div> | ||
+ | <h2 class="subtitle" | ||
+ | style="margin-top: 25px;">Düsseldorf meetup</h2> | ||
+ | <div style="display: flex; flex-direction: row;"> | ||
+ | <p> | ||
+ | From the 5th till the 7th July we went to the HHU for the German meetup. Many lectures with interesting topics | ||
+ | were offered by the Cluster of Exellence of Plant Science (<a href="https://www.ceplas.eu/en/home/" | ||
+ | target="_blank">CEPLAS</a>) by Prof. Andreas Weber | ||
+ | but also non-scientific aspects like courses on presentations. We tried to visit all of | ||
+ | them | ||
+ | while presenting our poster in the hall. There we got in touch with other iGEM Teams and also | ||
+ | learnt | ||
+ | about their projects. We promoted our Webinar project and laid the foundation stone for | ||
+ | future | ||
+ | collaborations. On the second day, iGEM Düsseldorf organized a panel discussion about the | ||
+ | issues | ||
+ | concerning green genetic engineering. Thereby, we got great input on how to structure our | ||
+ | panel | ||
+ | discussion and which subjects should be mentioned. Beside the regular program organized by | ||
+ | iGEM | ||
+ | Düsseldorf we used a lot of chances to get in contact with other teams. After the | ||
+ | presentations | ||
+ | a couple of tourist guides showed us the beautiful old town of Düsseldorf and explained | ||
+ | some | ||
+ | interesting historical facts. Afterwards we had again the possibility to meet with other | ||
+ | iGEMers | ||
+ | in a pub and talk about the projects, especially to find weaknesses or opportunities for | ||
+ | collaborations. This really went successfully due to the time we spend with iGEM Team Aachen, Kaiserslautern and Hamburg. The iGEM Team Düsseldorf also arranged a game contest between the teams, | ||
+ | where | ||
+ | we lost in the final to our good friends from Potsdam. Congrats! <br> | ||
+ | Thanks again to iGEM Düsseldorf for the huge effort and the great meetup! | ||
+ | </p> | ||
+ | <figure style="text-align:center; margin-left: 25px;"> | ||
+ | <img style="min-width: 200px;" | ||
+ | src="https://static.igem.org/mediawiki/2019/e/e4/T--marburg--meet_up_Duesseldorf.jpeg" | ||
+ | alt="Du meetup"> | ||
+ | <figcaption> | ||
+ | Fig. 3: Team photo from the Düsseldorf meetup. | ||
+ | </figcaption> | ||
+ | </figure> | ||
+ | </div> | ||
+ | <h2 class="subtitle" | ||
+ | style="margin-top: 25px;">Vilnius, Lithuania – Biohackathon Lab App 2019 | ||
+ | </h2> | ||
+ | <div style="display: flex; flex-direction: row;"> | ||
+ | |||
+ | <figure style="text-align:center; margin-right: 25px"> | ||
+ | <img style="min-width: 300px;" | ||
+ | src="https://static.igem.org/mediawiki/2019/4/4e/T--Marburg--vilnius.jpg" | ||
+ | alt="Vilnius biohackathon"> | ||
+ | <figcaption> | ||
+ | Fig. 4: Biohackathon in Vilnius. | ||
+ | </figcaption> | ||
+ | </figure> | ||
+ | <p> | ||
+ | In August 2019 our Team participated in the Biohackathon Lab App, organized by the | ||
+ | Vilnius-Lithuania Team 2019. Together with the iGEM Teams Stockholm and UCopenhagen, as | ||
+ | well | ||
+ | as | ||
+ | several non-iGEM Teams we sat together for a weekend packed with problems, ideas and neat | ||
+ | solutions. | ||
+ | With the task of creating an application that tackles a self-found problem, we sat | ||
+ | together | ||
+ | and | ||
+ | brainstormed about what could be done. | ||
+ | <br> | ||
+ | During this time we were starting to work more with the Opentrons OT-2 and soon realized | ||
+ | that | ||
+ | some | ||
+ | of our biologists had trouble integrating protocols in the robot themselves, as this is | ||
+ | primarily | ||
+ | done by writing the protocol in Python and pushing it directly to the robot – no | ||
+ | convenient | ||
+ | application to change parameters, such as the amount of reactions,was included in the | ||
+ | protocols. | ||
+ | In our heads the idea to have a user-friendly GUI grew more and more and soon we found | ||
+ | ourselves | ||
+ | standing in the Vilnius University Life Sciences Center. | ||
+ | <br>“We” were two biologists and one mathematician, so we talked to some of the other | ||
+ | participants | ||
+ | that did not have a team yet and found the perfect fit to complement our small team: Nour | ||
+ | Alsamsam | ||
+ | was happy to join us for this weekend and provided us with his coding skills – huge thank | ||
+ | you | ||
+ | to | ||
+ | you, Nour! | ||
+ | Together we tried our best to come up with a suitable app and presented it in the end, | ||
+ | having | ||
+ | more | ||
+ | ideas in our heads than we could implement in such a short time – it was clear for us, | ||
+ | that we | ||
+ | would continue with this project later on! | ||
+ | <br>In conclusion, the Biohackathon was an awesome and productive meeting of the iGEM | ||
+ | teams | ||
+ | Stockholm, Copenhagen, Vilnius and Marburg. We talked about our projects, offered each | ||
+ | other | ||
+ | feedback on current issues in the lab and shared many ideas. | ||
+ | We are very grateful that we got the opportunity to meet so many experts from different | ||
+ | fields, | ||
+ | which really helped getting a fresh perspective on our project! | ||
+ | </p> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
</div> | </div> | ||
+ | </section> | ||
</div> | </div> | ||
− | |||
− | |||
</html> | </html> | ||
{{Marburg/footer}} | {{Marburg/footer}} |
Latest revision as of 06:45, 9 December 2019
C O L L A B O R A T I O N S
Through this iGEM year we interacted with many iGEM Teams all around the world. It was a pleasure working with all the different teams and learn from each other. We got many impressions about other projects and we hope that we could help with our collaborations. We would like to thank every team that collaborated with us!
C O L O N Y
P I C T U R E S
The Colony Picture Collaboration
In our colony picking collaboration we asked other iGEM teams to supply us with data for our picking algorithm. Thanks to this group effort we could build our Colony Picking Unit.
![Pipetting Robot, colony picking set up](https://static.igem.org/mediawiki/2019/5/5e/T--marburg--colony_picking_2.jpeg)
Many repetitive tasks in the lab cost time such as pipetting or picking colonies from agar plates and could technically be performed by robots. However, most commercially available solutions are simply too expensive and therefore unavailable for many Labs and iGEM Teams. So one of our goals was to build a reasonable colony picking robot to outsource the tiring task of colony picking by hand.
We used an Opentrons OT-2, the cheapest available pipetting robot and constructed modules and scripts, that allow other iGEM Teams to turn their own OT-2 into a colony picking robot as well. See here how to transfer your OT2 into a Colony Picker.
We chose to build an Artificial Intelligence (AI) to automatically recognise colonies. While a “hard-coded” solution would be easier to achieve, they lack accuracy and flexibility, key attributes of Synbio projects. The drawback for an AI approach is the amount of data it needs for precise and robust training. With the help of the iGEM-community we thought to overcome this problem. We asked other iGEM Teams if they could provide us with labeled pictures of their agar plates with E. coli colonies, which served as the basis for our AI. For that purpose we set up a Website.
To participate all that was needed were agar plates with E. coli colonies, a screen and a camera. Other criteria were evenly distributed, distinguishable colonies. The first step was taking the picture. For this purpose, the screen had to be set to the highest level of brightness and display a white image. When you take a picture of a common screen, a pattern on the picture could appear, the so called Moiré pattern. To avoid this, we supplied each team with diffusive sheets, which they could use to cover their screen. A detailed summary is found here.
![Pattern for taking the colony picture](https://static.igem.org/mediawiki/2019/f/f8/T--marburg--taking_colony_pictures.png)
![colony picking website](https://static.igem.org/mediawiki/2019/0/0b/T--marburg--collonycollab_website.png)
Following this, the images taken had to be labeled using a web-based tool. Afterwards, the single colonies were marked with a rectangle. In total 250 colony pictures were added to our data set, vastly increasing the performance of our AI in detecting colonies. As an incentive to participate in this collaboration we gave away prizes to the teams that sent us their pictures.
As an incentive to participate in this Collaboration we gave away prizes. The five best teams that sent us pictures got an E. coli plushie and the three teams that sent us the most pictures got a bigger surprise. Thanks to Doulix, we provided these three teams with an engraved trophy out of glass and vouchers.
Team BOKU-Vienna was able to send us one picture, team GO Paris-Saclay provided three pictures and team TUDelft could help us with 14 colony pictures. Team TU Darmstadt could send us a number of 23 pictures, but were beaten by team IISER Bhopal which send us an impressive amount of 99 pictures.
![Prizes colony picture](https://static.igem.org/mediawiki/2019/0/04/T--Marburg--prizes_colony-picture.jpeg)
We thank all the teams for sending us labeled colony pictures. Without your help we would not have been able to train our AI and to transform our Opentrons OT-2 into a Colony Picker.
T H E G O L D E N G A T E
C O L L A B O R A T I O N
The Golden Gate collaboration
Our team made itself the goal to introduce other iGEM Teams to Golden Gate. Therefore we hosted a webinar, build up a communication platform and made an Interlab study.
Earlier this year the iGEM headquarters announced changes in their part submission system, one of
which made us extremely happy: BioBricks are now also accepted in Type IIS standard! In our opinion,
this is a huge step, as cloning techniques based on Type IIS restriction enzymes are becoming more
and more popular in the scientific community due to their numerous advantages over classical cloning
methods.
We noticed that more teams converted to MoClo systems in their projects this year but we still think
there is a lot that needs to be done for the iGEM community to really embrace this change. In order
to help teams with the transition into this new system, we had an idea:
What if we could help other teams understand what Type IIS enzymes really are and how they work? If
we could share our passion for Modular Cloning and the value of the PhytoBrick standard? If we could
introduce them to our Marburg Collection and help removing some of the burdens that come with a
switch to a new cloning system?
Therefore we hosted a webinar about Golden Gate, build up a communication platform to give iGEM
Teams a chance to troubleshoot their methods and made an Interlab study. Here we evaluated the
consistency of Golden Gate Assembly protocols across iGEM teams as well as to give teams the
opportunity to gain experience with the assembly method.
We wanted to reach as many teams as possible with our webinar on Golden Gate cloning,
Type IIS enzymes, their benefits and how to use all of this in their own projects. We
started planning everything that we thought should be in the webinar, from the
biological function of the Type IIS enzymes and how they differ from Type II enzymes,
over its applications in Synthetic Biology like modular cloning to the in-depth
strategies when designing your own parts and actually building them in the lab.
During this planning step we realized, that some of the teams that are already working
with Golden Gate might not benefit as much from an entry level webinar. Consequently, we
thought about integrating more complex design questions that we could share with the
community. We finished our webinars with a practical part, showing the part-design in
Geneious Prime – the sequence analysis software we used throughout the year. Firstly, we
explained the different levels that can be built with Golden Gate and then introduced
some more complex designs that are also possible with Golden Gate cloning, such as the
construction of whole operons or space holder parts. Through this we hoped to also reach
teams that use Golden Gate themselves, showing them the possibilities this cloning
system offers.
To reach as many teams on the globe as possible we set up webinars on three different times to cover as many time zones within a comfortable time frame – hoping that this will allow more iGEM teams to participate. The one and a half hour webinars were held on July 30th 10am CET, July 31st 6pm CET and the last time on 8th August 12pm CET by our instructor René Inckemann with the support of three of our team members. We were happy that in total 18 teams participated, as well as some students from the universities of Marburg and Gießen and even a few fellow scientists from the Max-Planck-Institute for terrestrial microbiology. In total we counted around 70 participants. We were also really happy to see Sota Hirano from Doulix on our webinar, who gave us some awesome feedback, stating that he did learn new things about Golden Gate which he was not aware of before!
![Bilder Webinar](https://static.igem.org/mediawiki/2019/f/f1/T--marburg--Doulix_webinar.jpeg)
If you would like to learn about Golden Gate assembly please feel free to watch our webinar through this Link:
We thank all the following teams for participating in this awesome event:
Although we were quite pleased with the result of our webinar, we thought about how to
further connect this Golden Gate community, so we created a Slack workspace for everyone
that is working with this method, enabling us all to exchange protocols and ideas or
troubleshoot issues that might arise. So far there are close to 50 members, but we hope
to find more enthusiasts willing to share their knowledge!
All files used for the webinar are openly available. You can find our protocols, handouts, slides as well as the files we used for the practical part in Geneious to construct parts in-silico in our shared Google Drive. Through this we hope to make it easier for other teams willing to start with Golden Gate cloning, as they can start designing their first own in-silico parts with our guidance.
The main documents of the Golgen Gate Webinar
→ Download: Golden
Gate webinar slide 2019
→ Download: Handout
Golden Gate webinar
→ Download: Golden
Gate protocoll
To try out the described in-silico cloning you can download the needed sequences as
geneious file here.
→ Download: Sequences
Webinar
As we are constantly working on improving the efficiency of Golden Gate. We clearly
saw the main bottleneck of the reaction was the long assembly time in the cycler.
In the webinar and in our Golden Gate community we provided protocols with three different
cycler settings for different applications.
Although these three protocols were tested before, they were never compared to each other in
their efficiency.
We conducted our own Interlab study in collaboration with the iGEM Teams from
TUDelft ,
HUmboldt Berlin
and
TU Kaiserslautern
In order to find the best protocol, we supplied the teams with specific Level 0
parts,that should be assembled to a Level 1 vector using the three different cycler
conditions. The provided parts are shown in the table
5' Connector | BBa_K2560075 / |
Promoter | J23119 BBa_K2560031 |
RBS | B0034 BBa_K2560008 |
CDS | sfGFP BBa_K2560042 |
Terminator | B0015 BBa_K2560035 |
3' Connector | BBa_K2560080 |
ORI: | ColE1 BBa_K2560036 |
Antibiotic-Resistance-Cassette | Kan-RFP BBa_K2560056 |
![Content of the Interlab package](https://static.igem.org/mediawiki/2019/9/91/T--marburg--interlab_package.jpeg)
The Golden Gate reactions should be pipetted together using the given parts and then
be transformed in E. coli and be plated with similar amounts on agar plates. A colony with a
correct assembled
vector would have a functioning transcription unit which expresses GFP, which can be clearly
distinguished from RFP expressing colonies which only carries the antibiotic cassette and
false non fluorescent
colonies.
The ratio of green to non-green colonies was used to evaluate which of the three protocols
yielded in the
highest transformation efficiency and the highest rate of positive clones.
The handout with the used protocols can be downloaded here .
Results
First we wanted to know which protocol worked best for every team. For this we analysed the data of the correct assembled green colonies for every protocol. The data of the different teams are given in Table 1 and are shown in as chart in Figure 2. The total colony count is hard to compare directly, cause the numbers differ very much between the teams. This could come through the different transformation efficiencies for example because of the variating competent cells the teams used.
![Table 1 - Data for the gained green colonies gained by using the three different protocols, from each participating team](https://static.igem.org/mediawiki/2019/4/4a/T--Marburg--Tabelle_Interlab.png)
![Amount of gained green colonies gained by using the three different protocols, from each participating team.](https://static.igem.org/mediawiki/2019/3/3a/T--Marburg--Interlab_1.png)
The results in Figure 2 show, that the protocols work differently for each team. In general the most colonies were
received with the long Troubleshoot protocol, but also the Rapid and Improved protocols show high numbers of correct colonies.
But which protocol is the most efficient one?
To compare the results of the teams the ratio of the gained correct colonies between the
protocols were averaged. The data are shown in Figure 3 and show, that the long Troubleshoot
assembly works the best. The fastest assembly (Rapid) works the second best. This is very
interesting, because this assembly takes significant less time.
We would propose to take the Troubleshoot protocol for complicated assemblies and the rapid
protocol for easy assemblies like lvl 0.
![Fig.3 Average efficiency of the three protocols](https://static.igem.org/mediawiki/2019/a/a1/T--Marburg--Interlab_3.png)
B E R L I N
![](https://static.igem.org/mediawiki/2019/9/9b/T--marburg--berlin_logo_richtig.png)
Berlin Collaboration
iGEM Berlin engineered the freshwater algae Chlamydomonas reinhardtii to degrade
Polyethylene
terephthalate
(PET). For this goal they also built a bioreactor for their chassis. We sent
them our UTEX
strain to test if
their photobioreactor is capable of cultivating other phototrophic organisms as
well. They gained
additional
experiences on growth conditions for phototrophic organisms and we could see our
strains grow in
a bioreactor environment.
Click here to learn more about there project Chlamylicious
![Berlin logo](https://static.igem.org/mediawiki/2019/9/9b/T--marburg--berlin_logo_richtig.png)
D E L F T
Delft Collaboration
![DELFT Bild](https://static.igem.org/mediawiki/2019/5/5e/T--marburg--Skype_DELF.jpeg)
Delft Collaborations
iGEM Delft created an all species encompassing toolbox, based on orthogonal gene
expression and
host
independent plasmid replication. We exchanged multiple times over Molecular
Cloning and toolbox
designs.
Additionally, we sent them parts in the PhytoBricks standard.
Click here to learn more about there project
project SCI-PHI 29
U C D A V I S
![](https://static.igem.org/mediawiki/2019/d/d7/T--UC_Davis--Teamlogolight.png)
UC Davis collaboration
UC Davis iGEM team is one of the winners of the Opentrons competition in 2019. iGEM Marburg won one OT-2 during last year’s edition of the competition and got contacted for a collaboration by iGEM UC Davis along with other winners from 2018 and 2019. The goal for this automation InterLab collaboration was to collect data to determine the accuracy and reproducibility of the OT-2 compared to humans. To do this, UC Davis proposed to perform the standard iGEM protocol on serial dilutions with silica beads and fluorescein, each manually and with an OT-2. In total iGEM Marburg did four dilutions; twice with an OT-2 and twice manually. The data was then sent to UC Davis via the spreadsheet designed by them. Finally, iGEM Marburg aided by creating a Slack channel for all iGEM teams with an OT-2 to facilitate better communication and exchange in the future.
![UCD OT2](https://static.igem.org/mediawiki/2019/4/49/T--marburg--UCD_OT2.jpeg)
In total, seven teams participated and sent their data to UC Davis which are
shown in the
figure below.
According to UC
Davis, the fluorescence data was standardized using the iGEM MEFL conversion to
standardize
the plate reader
measurements. Subsequently, the data was fitted using a linear regression model,
whose
quality is quantified
using the
r2 metric (it ranges from 0 to 1 with higher value indicates a better fit).
For the fluorescence the r2 value from the OT2 data beat human r2 value with
0.94 and 0.90,
respectively.
Interestingly, for the silica beads, manual measurements yielded a more
satisfactory result
with r2 value of
0.90
compared to 0.82 from OT2 measurements. One possible explanation could be the
speed of the
OT-2: it took
twice as long
to complete this task with the OT-2 compared to a manual trial. This could lead
to the
sedimentation of the
silica beads
as the OT-2 went through their runs; therefore worsening its results. Also, as
shown in Fig.
2b, one team’s
result
deviates a lot from the rest of the teams, maybe if their result is taken out from
the overall
evaluation a more
comparable
result between OT-2 and human could be observed.
More information about this collaboration such as individual team data can be found on UC Davis wiki.
![UCD Graphic 1](https://static.igem.org/mediawiki/2019/c/cc/T--marburg--UC_Davis_Graphic_1.png)
![UC Davis Graphic 2]( https://static.igem.org/mediawiki/2019/a/a7/T--marburg--UC_Davis_Graphic_2.png)
U F L O R I D A
![](https://static.igem.org/mediawiki/2019/f/f7/T--Florida--iGEMWeb1.png)
UFlorida
![Florida](https://static.igem.org/mediawiki/2019/f/f7/T--Florida--iGEMWeb1.png)
This semester we had the pleasure of mentoring UFlorida for their Wiki
design. Our member, René Inckemann, aided UFlorida in their quest to create
a visually accessible Wiki, similar to what we did this past iGEM season.
We helped shape UFlorida’s website into an interactive and appealing
landscape, while maintaining the integrity of their goal and project. We
advised them on their font choices- specifically their use of bold and
italicized words-, along with answering questions about our work with the
visually disabled community and utilizing our “Accessible Wiki Guide” from
the previous year.
Besides answering questions specifically about their project, we also
offered valuable information about when to start working on the Wiki and
questions about coding. Working with a young team like UFlorida offered us
a great mentorship opportunity.
Click here to learn more about there project.
![Florida](https://static.igem.org/mediawiki/2019/2/29/T--marburg--UFlorida_Rene.jpeg)
I N T E R A C T I O N S
W I T H
O T H E R T E A M S
![](https://static.igem.org/mediawiki/2019/f/fe/T--marburg--Postcard.png)
Interactions with other teams
Mike the microbe
![Mike the Mircrobe](https://static.igem.org/mediawiki/2019/b/b0/T--marburg--Mike_microbe.png)
We participated in the “Mike the microbe” collaboration of the iGEM Team US AFRL CarrollHS. They sent us their virtual mascots Mike the Microbe and Chia the Chitinase so we could take pictures with them to connect all iGEM teams around the world via social media. It brought us great joy to join to this collaboration and let the two little buddies visit our daily life in our incubators for cyanobacteria.
Postcards Düsseldorf
iGEM Düsseldorf called for their traditional collaboration project, a long year postcard campaign with the purpose to educate and to show the public about the different topics and projects in Synthetic Biology and especially regarding all the iGEM projects. With interesting designs and a short project abstract iGEM teams could spread their ideas using the postcards to share their thinking to each other and the society. Our title and postcard design was about our photosynthetic project „Syntex“. You can see Synechococcus elongatus UTEX 2973 direly needing a refreshing drink, a cute little draft from our team member Jonas.
![Postcard challenge](https://static.igem.org/mediawiki/2019/f/fe/T--marburg--Postcard.png)
iGEMxSDGs challenge
![SGD Challenge - 7 12 13 15](https://static.igem.org/mediawiki/2019/1/18/T--marburg--SDG_chalenge.png)
IGEM Costa Rica, Tuebingen, and TAS Taipei created an iGEMxSDGs challenge as collaboration. They asked us to choose four of the Sustainable Development Goals (SDGs) proposed by the United Nations (UN). We joined this collaboration challenge and spread it on our social media account. Our project targets the goals shown in Figure 3.
Who wore it best? #labcoatchallenge
The Stony Brook iGEM Team challenged us to design an outfit for a runway from a standard lab coat. The outfit had to be created by tying, not using duct tape, cut or sew, etc. After designing our outfits four of our members presented their style on our ‘runway’ while paparazzi took their photos. As an outfit we chose a dress highlighting the neckline, a shoulder free dress, a knotted dress and a skirt to bring glamour to the daily routine in the labs. Especially our guys were rocking their outfits as you can see in Figure 4.
![Our model Burgis for the labcoatchallenge](https://static.igem.org/mediawiki/2019/7/74/T--marburg--Labcoat_Burgi.jpeg)
M E E T U P S
![](https://static.igem.org/mediawiki/2019/9/90/T--Marburg--meetups.jpg)
Meet ups
iGEM Spring Festival in Bonn
From the 3rd to the 5th Mai we met other iGEMers at the Spring Festival in Bonn. Professors
of the university of Bonn presented their research fields which was met with huge interest. We heard
lectures
about immunology and neurosciences, as well as reviving mammoths. During the lectures,
specific
topics were deepened in small seminar groups, such as the ethical tenability of CRISPR
applied
at humans and other animals. At the poster sessions we got the first time in contact with
other teams’ projects. Afterwards, we had the opportunity to vote for the best project,
which
iGEM Düsseldorf won. Besides that, the meetup was filled with a lot of awarding
categories.
We
competed in the categories for “Best Video”, “Best Poster”, “Best Project” and a single
person
had the chance to win the award for “Best Meme”. The iGEM Team Bonn offered spontaneously
a
little football table tournament, where two of our team members won. In the evening we
learnt a
lot about Bonn as a city from a guide living there for many years. Subsequently, the iGEM Teams
met
in
several groups in restaurants and bars. We merged with iGEM Kaiserslautern and exchanged
our
projects and ideas for the rest of the evening.
On the second day, the iGEM Team Bonn offered a competition out of three rounds, where we
competed against other teams. We were asked iGEM specific questions, had to solve riddles
for an
escape room and finished a lab challenge afterwards. Our team could compete until the end
and
won the prize! Thanks again to Doulix for this awesome prize!
We particularly thank the iGEM Team Bonn for the huge effort and the great meetup!
![Erlangen meetup](https://static.igem.org/mediawiki/2019/a/a9/T--marburg--meet_u_bonn.png)
Erlangen collaboration meetup
![Erlangen meetup](https://static.igem.org/mediawiki/2019/0/01/T--marburg--meet_up_erlangen.jpeg)
On the 17th till the 19th of May we attended the collaboration meetup hosted by the iGEM team Erlangen 2019. There, we met the iGEM teams of Straßbourg, Tübingen, Vienna and of course Erlangen. Apart from listening to the interesting talks from the various professors there, we made a lot of new friends and took the chance to talk about possible collaborations with the other teams and also promote our collaborations, such as the colony picking project and the Golden Gate webinar. We would like to thank iGEM team Erlangen very much for organizing this wonderful meetup!
Düsseldorf meetup
From the 5th till the 7th July we went to the HHU for the German meetup. Many lectures with interesting topics
were offered by the Cluster of Exellence of Plant Science (CEPLAS) by Prof. Andreas Weber
but also non-scientific aspects like courses on presentations. We tried to visit all of
them
while presenting our poster in the hall. There we got in touch with other iGEM Teams and also
learnt
about their projects. We promoted our Webinar project and laid the foundation stone for
future
collaborations. On the second day, iGEM Düsseldorf organized a panel discussion about the
issues
concerning green genetic engineering. Thereby, we got great input on how to structure our
panel
discussion and which subjects should be mentioned. Beside the regular program organized by
iGEM
Düsseldorf we used a lot of chances to get in contact with other teams. After the
presentations
a couple of tourist guides showed us the beautiful old town of Düsseldorf and explained
some
interesting historical facts. Afterwards we had again the possibility to meet with other
iGEMers
in a pub and talk about the projects, especially to find weaknesses or opportunities for
collaborations. This really went successfully due to the time we spend with iGEM Team Aachen, Kaiserslautern and Hamburg. The iGEM Team Düsseldorf also arranged a game contest between the teams,
where
we lost in the final to our good friends from Potsdam. Congrats!
Thanks again to iGEM Düsseldorf for the huge effort and the great meetup!
![Du meetup](https://static.igem.org/mediawiki/2019/e/e4/T--marburg--meet_up_Duesseldorf.jpeg)
Vilnius, Lithuania – Biohackathon Lab App 2019
![Vilnius biohackathon](https://static.igem.org/mediawiki/2019/4/4e/T--Marburg--vilnius.jpg)
In August 2019 our Team participated in the Biohackathon Lab App, organized by the
Vilnius-Lithuania Team 2019. Together with the iGEM Teams Stockholm and UCopenhagen, as
well
as
several non-iGEM Teams we sat together for a weekend packed with problems, ideas and neat
solutions.
With the task of creating an application that tackles a self-found problem, we sat
together
and
brainstormed about what could be done.
During this time we were starting to work more with the Opentrons OT-2 and soon realized
that
some
of our biologists had trouble integrating protocols in the robot themselves, as this is
primarily
done by writing the protocol in Python and pushing it directly to the robot – no
convenient
application to change parameters, such as the amount of reactions,was included in the
protocols.
In our heads the idea to have a user-friendly GUI grew more and more and soon we found
ourselves
standing in the Vilnius University Life Sciences Center.
“We” were two biologists and one mathematician, so we talked to some of the other
participants
that did not have a team yet and found the perfect fit to complement our small team: Nour
Alsamsam
was happy to join us for this weekend and provided us with his coding skills – huge thank
you
to
you, Nour!
Together we tried our best to come up with a suitable app and presented it in the end,
having
more
ideas in our heads than we could implement in such a short time – it was clear for us,
that we
would continue with this project later on!
In conclusion, the Biohackathon was an awesome and productive meeting of the iGEM
teams
Stockholm, Copenhagen, Vilnius and Marburg. We talked about our projects, offered each
other
feedback on current issues in the lab and shared many ideas.
We are very grateful that we got the opportunity to meet so many experts from different
fields,
which really helped getting a fresh perspective on our project!