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| | margin-left: 30%; | | margin-left: 30%; |
| | margin-top: 15%; | | margin-top: 15%; |
| − | position: center;
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| | } | | } |
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| | .offset-xl-11 { | | .offset-xl-11 { |
| | margin-left: 91.666667% | | margin-left: 91.666667% |
| | + | } |
| | + | } |
| | + | |
| | + | .veggie { |
| | + | width: 100%; |
| | + | max-width: 100%; |
| | + | margin-bottom: 1rem; |
| | + | background-color: transparent; |
| | + | z-index:100; |
| | + | } |
| | + | .veggie td, |
| | + | .veggie th { |
| | + | padding: .75rem; |
| | + | vertical-align: top; |
| | + | border-top: 1px solid #F4F0DE; |
| | + | color: #F4F0DE; |
| | + | border-bottom: 1px solid #F4F0DE; |
| | + | z-index:100; |
| | + | } |
| | + | .veggie th { |
| | + | background-color: #2ecc71 !important; |
| | + | color: #232323 !important; |
| | + | z-index:100; |
| | + | } |
| | + | .veggie caption { |
| | + | color: #adb3b7; |
| | + | z-index:100; |
| | + | } |
| | + | .veggie thead th { |
| | + | vertical-align: bottom; |
| | + | border-bottom: 2px solid #dee2e6; |
| | + | z-index:100; |
| | + | } |
| | + | .veggie tbody+tbody { |
| | + | border-top: 2px solid #dee2e6; |
| | + | z-index:100; |
| | + | } |
| | + | |
| | + | @media screen and (max-width: 800px) { |
| | + | .veggie { |
| | + | font-size: xx-small; |
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| | display: block; | | display: block; |
| | width: 100%; | | width: 100%; |
| − | overflow-x: auto; | + | overflow-x: break-words; |
| | -webkit-overflow-scrolling: touch; | | -webkit-overflow-scrolling: touch; |
| | -ms-overflow-style: -ms-autohiding-scrollbar | | -ms-overflow-style: -ms-autohiding-scrollbar |
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| | display: block; | | display: block; |
| | width: 100%; | | width: 100%; |
| − | overflow-x: auto; | + | overflow-x: break-words; |
| | -webkit-overflow-scrolling: touch; | | -webkit-overflow-scrolling: touch; |
| | -ms-overflow-style: -ms-autohiding-scrollbar | | -ms-overflow-style: -ms-autohiding-scrollbar |
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| | display: block; | | display: block; |
| | width: 100%; | | width: 100%; |
| − | overflow-x: auto; | + | overflow-x: break-words; |
| | -webkit-overflow-scrolling: touch; | | -webkit-overflow-scrolling: touch; |
| | -ms-overflow-style: -ms-autohiding-scrollbar | | -ms-overflow-style: -ms-autohiding-scrollbar |
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| | .bg-dark { | | .bg-dark { |
| − | background-color: #343a40 !important | + | background-color: #232323 !important |
| | } | | } |
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| | .dropdown-menu { | | .dropdown-menu { |
| − | background-color: rgba(100, 100, 100, 1); | + | background-color: #232323; |
| | } | | } |
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| | <li id="ourStoryButton" class="nav-item mx-3"> | | <li id="ourStoryButton" class="nav-item mx-3"> |
| | <a class="nav-link" href="/Team:Tuebingen/Story" | | <a class="nav-link" href="/Team:Tuebingen/Story" |
| − | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">OUR STORY</a> | + | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">OUR |
| | + | STORY</a> |
| | </li> | | </li> |
| | | | |
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| | <a class="dropdown-item" href="/Team:Tuebingen/Incretin" | | <a class="dropdown-item" href="/Team:Tuebingen/Incretin" |
| | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Incretin</a> | | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Incretin</a> |
| − | <a class="dropdown-item" href="/Team:Tuebingen/CPP" | + | <a class="dropdown-item" href="/Team:Tuebingen/Model" |
| − | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Cell Penetrating Peptides</a> | + | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Model</a> |
| | + | <a class="dropdown-item" href="/Team:Tuebingen/Software" |
| | + | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Software</a> |
| | <a class="dropdown-item" href="/Team:Tuebingen/Results" | | <a class="dropdown-item" href="/Team:Tuebingen/Results" |
| | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Results</a> | | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Results</a> |
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| | <a class="dropdown-item" href="/Team:Tuebingen/Notebook" | | <a class="dropdown-item" href="/Team:Tuebingen/Notebook" |
| | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Notebook</a> | | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Notebook</a> |
| − | <a class="dropdown-item" href="/Team:Tuebingen/Model"
| |
| − | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Model</a>
| |
| − | <a class="dropdown-item" href="/Team:Tuebingen/Software"
| |
| − | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Software</a>
| |
| | <a class="dropdown-item" href="/Team:Tuebingen/Outlook" | | <a class="dropdown-item" href="/Team:Tuebingen/Outlook" |
| | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Outlook</a> | | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Outlook</a> |
| | + | <a class="dropdown-item" href="/Team:Tuebingen/Entrepreneurship" |
| | + | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Entrepreneurship</a> |
| | </div> | | </div> |
| | </li> | | </li> |
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| | <div class="dropdown-menu"> | | <div class="dropdown-menu"> |
| | <a class="dropdown-item" href="/Team:Tuebingen/Parts" | | <a class="dropdown-item" href="/Team:Tuebingen/Parts" |
| − | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Parts Overview</a> | + | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Parts |
| | + | Overview</a> |
| | <a class="dropdown-item" href="/Team:Tuebingen/Basic_Part" | | <a class="dropdown-item" href="/Team:Tuebingen/Basic_Part" |
| − | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Basic Parts</a> | + | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Basic |
| | + | Parts</a> |
| | <a class="dropdown-item" href="/Team:Tuebingen/Improved_Part" | | <a class="dropdown-item" href="/Team:Tuebingen/Improved_Part" |
| − | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Improved Parts</a> | + | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Improved |
| | + | Parts</a> |
| | + | <a class="dropdown-item" href="/Team:Tuebingen/Characterized_Part" |
| | + | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Improved |
| | + | Parts</a> |
| | <a class="dropdown-item" href="/Team:Tuebingen/Composite_Part" | | <a class="dropdown-item" href="/Team:Tuebingen/Composite_Part" |
| − | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Composite Parts</a> | + | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Composite |
| | + | Parts</a> |
| | <a class="dropdown-item" href="/Team:Tuebingen/Part_Collection" | | <a class="dropdown-item" href="/Team:Tuebingen/Part_Collection" |
| − | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Part Collection</a> | + | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Part |
| | + | Collection</a> |
| | + | <a class="dropdown-item" href="/Team:Tuebingen/Downloads" |
| | + | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Downloads</a> |
| | </div> | | </div> |
| | </li> | | </li> |
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| | <a class="dropdown-item" href="/Team:Tuebingen/Collaborations" | | <a class="dropdown-item" href="/Team:Tuebingen/Collaborations" |
| | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Collaborations</a> | | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Collaborations</a> |
| − | <a class="dropdown-item" href="/Team:Tuebingen/Attributions"
| |
| − | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Attributions</a>
| |
| | <a class="dropdown-item" href="/Team:Tuebingen/Sponsors" | | <a class="dropdown-item" href="/Team:Tuebingen/Sponsors" |
| | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Sponsors</a> | | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Sponsors</a> |
| | + | <a class="dropdown-item" href="/Team:Tuebingen/Attributions" |
| | + | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Attributions</a> |
| | <a class="dropdown-item" href="/Team:Tuebingen/Gallery" | | <a class="dropdown-item" href="/Team:Tuebingen/Gallery" |
| | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Gallery</a> | | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Gallery</a> |
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| | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Overview</a> | | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Overview</a> |
| | <a class="dropdown-item" href="/Team:Tuebingen/Public_Engagement" | | <a class="dropdown-item" href="/Team:Tuebingen/Public_Engagement" |
| − | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Education & Public | + | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Education & Public Engagement</a> |
| − | Engagement</a>
| + | |
| | <a class="dropdown-item" href="/Team:Tuebingen/Experts" | | <a class="dropdown-item" href="/Team:Tuebingen/Experts" |
| | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Experts</a> | | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Experts</a> |
| − | <a class="dropdown-item" href="/Team:Tuebingen/Entrepreneurship"
| |
| − | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Entrepreneurship</a>
| |
| | <a class="dropdown-item" href="/Team:Tuebingen/SDG" | | <a class="dropdown-item" href="/Team:Tuebingen/SDG" |
| − | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Sustainable Development | + | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Sustainable Development Goals</a> |
| − | Goals</a>
| + | |
| | <a class="dropdown-item" href="/Team:Tuebingen/Experimenta" | | <a class="dropdown-item" href="/Team:Tuebingen/Experimenta" |
| − | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Experimenta Science | + | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Experimenta |
| − | Center</a> | + | Science Center</a> |
| | + | <a class="dropdown-item" href="/Team:Tuebingen/Survey" |
| | + | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Survey</a> |
| | </div> | | </div> |
| | </li> | | </li> |
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| | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Integrated Human Practise</a> | | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Integrated Human Practise</a> |
| | <a class="dropdown-item" href="/Team:Tuebingen/Public_Engagement" | | <a class="dropdown-item" href="/Team:Tuebingen/Public_Engagement" |
| − | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Education & Public | + | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Education & Public Engagement</a> |
| − | Engagement</a>
| + | |
| | <a class="dropdown-item" href="/Team:Tuebingen/Entrepreneurship" | | <a class="dropdown-item" href="/Team:Tuebingen/Entrepreneurship" |
| − | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Supporting | + | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Supporting Entrepreneurship</a> |
| − | Entrepreneurship</a>
| + | |
| − | <a class="dropdown-item" href="/Team:Tuebingen/Measurement"
| + | |
| − | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Measurement</a>
| + | |
| | <a class="dropdown-item" href="/Team:Tuebingen/Model" | | <a class="dropdown-item" href="/Team:Tuebingen/Model" |
| | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Model</a> | | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">Model</a> |
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| | <li class="nav-item mx-3"> | | <li class="nav-item mx-3"> |
| | <a class="nav-link" href="https://igem.org/2019_Judging_Form?team=Tuebingen" target="_blank" | | <a class="nav-link" href="https://igem.org/2019_Judging_Form?team=Tuebingen" target="_blank" |
| − | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">JUDGING | + | style="font-family: 'Righteous', cursive; font-weight: normal; color: #2ecc71;">JUDGING FORM ⇗</a> |
| − | FORM ⇗</a>
| + | |
| | </li> | | </li> |
| | | | |
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| | -webkit-transform: translateZ(0) scale(1.0, 1.0); | | -webkit-transform: translateZ(0) scale(1.0, 1.0); |
| | transform: translateZ(0); | | transform: translateZ(0); |
| − | background: #1B2030 url(https://2019.igem.org/wiki/images/a/a0/T--Tuebingen--description-banner.jpg) center no-repeat; | + | background: #1B2030 url(https://2019.igem.org/wiki/images/5/50/T--Tuebingen--experimenta-banner.jpg) center no-repeat; |
| | background-size: 100%, 100%; | | background-size: 100%, 100%; |
| | animation: grow 30s linear 10ms infinite; | | animation: grow 30s linear 10ms infinite; |
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| | } | | } |
| | | | |
| − | .header {
| |
| − | height:40vh
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| − | }
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| − | }
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| | | | |
| − | @media only screen and (max-width: 600px) {
| |
| − | .header {
| |
| − | height:35vh
| |
| − | }
| |
| | } | | } |
| | + | |
| | + | |
| | | | |
| | @media only screen and (max-width: 560px) { | | @media only screen and (max-width: 560px) { |
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| | margin-top: 15%; | | margin-top: 15%; |
| | } | | } |
| − | }
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| − |
| |
| − | @media only screen and (max-width: 500px) {
| |
| | .header { | | .header { |
| − | height:30vh | + | height: 40vh; |
| | } | | } |
| | } | | } |
| | + | |
| | + | |
| | | | |
| | @media only screen and (max-width: 470px) { | | @media only screen and (max-width: 470px) { |
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| | } | | } |
| | | | |
| − | @media only screen and (max-width: 450px) {
| + | |
| − | .header {
| + | |
| − | height:25vh
| + | |
| − | }
| + | |
| − | }
| + | |
| | | | |
| | @media only screen and (max-width: 400px) { | | @media only screen and (max-width: 400px) { |
| | .info h1 { | | .info h1 { |
| − | margin-top: 15%; | + | margin-top: 20%; |
| | + | } |
| | + | .header { |
| | + | height: 27vh; |
| | } | | } |
| | } | | } |
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| | @media only screen and (max-width: 350px) { | | @media only screen and (max-width: 350px) { |
| | .info h1 { | | .info h1 { |
| − | margin-top: 18%; | + | margin-top: 25%; |
| | } | | } |
| | } | | } |
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| | <div class="header"> | | <div class="header"> |
| | <div class="info"> | | <div class="info"> |
| − | <h1 style="font-family: 'Righteous', cursive; font-weight: normal; color:#2ecc71;">Notebook</h1> | + | <h1 style="font-family: 'Righteous', cursive; font-weight: normal; color:#2ecc71;">Experimenta</h1> |
| | </div> | | </div> |
| | </div> | | </div> |
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| | | | |
| − | <div style="background-color: #232323; overflow-x: fragments; overflow-wrap: inherit"> | + | <div style="background-color: #232323; overflow-x: fragments; overflow-wrap: break-word"> |
| | | | |
| | <div class="glpContainer"> | | <div class="glpContainer"> |
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| | <div id="image-popups"> | | <div id="image-popups"> |
| | | | |
| − | <div class="container-fluid py-5 mb-0" style="background-color: #232323"> <!-- FOR REUSE: REMOVE bg-dark IF U DONT WANNA HAVE A DARK BACKGROUND -->
| + | <h2 style="font-family:'Righteous'; color:#2ecc71;">The Experimenta Science Center Heilbronn</h2> |
| − | <div id="accordion">
| + | <div class="row"> |
| − | <div class="card">
| + | <div class="col-12"> |
| − | <div class="card-header" id="headingOne">
| + | <p> |
| − | <h5 class="mb-0">
| + | The Experimenta Science Center (<a href="https://www.experimenta.science/" onclick='window.open("https://www.experimenta.science/","_self");'>Experimenta</a>) in Heilbronn, Baden-Wuerttemberg, |
| − | <button class="btn btn-one btn-link collapsed" data-toggle="collapse" data-target="#collapseOne" aria-expanded="false" aria-controls="collapseOne">
| + | Germany offers science at your fingertips on an area of around 25,000 square metres with around 275 |
| − | Week 1 (08 Jul - 12 Jul)
| + | interactive exhibits, four creative studios and nine laboratories, as well as an observatory and an |
| − | </button>
| + | extraordinary science dome with spectacular science shows. Additionally, last year, they set up the |
| − | </h5>
| + | “Schülerforschungszentrum” (SFZ), where young researchers are actively supported in their initial |
| − | </div>
| + | scientific work. It is directed at students from high schools in Heilbronn. Their general idea is to |
| − | <div id="collapseOne" class="collapse" aria-labelledby="headingOne" data-parent="#accordion" >
| + | encourage students to develop their own projects while funding, lab space and supervision is granted. |
| − | <div class="card-body" style="background-color: #232323;">
| + | </p> |
| − | <p style="color: #F4F0DE">
| + | </div> |
| − | 08 Jakob, Patrick, Eva After moving into our new lab and settling in, first practical
| + | </div> |
| − | steps like aliquoting our primers or preparing LB-Medium and LB-Agar were taken.
| + | |
| − | 09 Jakob, Patrick, Eva We amplified Cas 3 out of E.Coli gDNA via PCR (Q5 Polymerase)
| + | <h2 style="font-family:'Righteous'; color:#2ecc71;">Our 4 Day Lab Course</h2> |
| − | and transformed the biobricks BBa_K608351 (K2), BBa_K091001 (K5), BBa_I13453 (K10),
| + | <div class="row"> |
| − | BBa_K584000 (AraC Promotor), BBa_K117008 (LsrR Promotor), BBa_R0073 (Mnt Promotor) in
| + | <div class="col-12"> |
| − | competent E. Coli DH5-alpha (NEB iGEM Kit). 10 Eva, Patrick We amplified Cascade out of
| + | <p>The course was held in a collaborative manner over a span of four days. Dr. Robert Friedrich, |
| − | E.Coli MG1655 gDNA via PCR (Q5 Polymerase) and checked the Cas 3 and Cascade PCRs via agarose
| + | a biologist working for the SFZ, gave the general lab introduction and provided the overall lab |
| − | gel electrophoresis. (figure 1) We picked colonies for all but the Ba_K608351 (K2)
| + | responsibility, while we were mainly responsible to teach the required theoretical background, |
| − | transformations (no growth) for inoculating overnight cultures and made a new batch of
| + | introduce bioinformatics data analysis (DNA-Seq) and finally project development. Additionally, |
| − | competent E.Coli Dh-5α cells. 11 Jakob, Eva, Patrick By Miniprep (Qiagen) we extracted the
| + | we were supporting Dr. Friedrich during the experiments and closely supervised the students.</p> |
| − | DNA of the overnight cultures. Amplification via PCR (Q5 Polymerase) of Cascade was repeated
| + | </div> |
| − | (figure 2). BBa_K608351 (K2) was transformed again and the competent cells tested.
| + | </div> |
| − | </p>
| + | <div class="row"> |
| − | <figure>
| + | <div class="col-12"> |
| | + | <div class="row align-items-center"> |
| | + | <div class="col-12 col-md-6"> |
| | + | <figure> |
| | <a style="font-size: small" | | <a style="font-size: small" |
| − | href="https://2019.igem.org/wiki/images/a/a8/T--Tuebingen--Gel1.png" | + | href="https://2019.igem.org/wiki/images/e/e8/T--Tuebingen--Experimenta-Lab4.png" |
| | data-effect="mfp-zoom-out" | | data-effect="mfp-zoom-out" |
| | title="Some title"> | | title="Some title"> |
| − | <img src="https://2019.igem.org/wiki/images/a/a8/T--Tuebingen--Gel1.png" | + | <img src="https://2019.igem.org/wiki/images/e/e8/T--Tuebingen--Experimenta-Lab4.png" |
| | class="figure-img img-fluid rounded" | | class="figure-img img-fluid rounded" |
| − | alt="A generic square placeholder"></a> | + | alt="Placeholder"></a> |
| − | <figcaption style="color: #F4F0DE; font-size: small;">Figure 1. 17/10/19 Gelelctrophoresis of Cas 3 and Cascade PCR. Gel loaded (from left to right): | + | </figure> |
| − | Ladder 1kb, Cas 3 (72°C), Cas 3 (68°C), H2O, Cascade (72°C), Cascade (68°C), H2O. It shows positive
| + | |
| − | PCR of Cas 3 (68°C), expected at ~2.7 kb.</figcaption>
| + | </div> |
| − | </figure>
| + | <div class="col-12 col-md-6 "> |
| − | <figure>
| + | <p>We conveyed the required theoretical background in a modern and interactive way, with the use of |
| | + | smart boards and current web tools. We focused on teaching the highly motivated students cutting-edge |
| | + | research, while providing a theoretical foundation, preparing them for different biology competitions |
| | + | and eventually university.</p> |
| | + | </div> |
| | + | </div> |
| | + | </div> |
| | + | </div> |
| | + | |
| | + | <div class="row"> |
| | + | <div class="col-12"> |
| | + | <div class="row align-items-center"> |
| | + | <div class="col-12 col-md-6"> |
| | + | <figure> |
| | <a style="font-size: small" | | <a style="font-size: small" |
| − | href="https://2019.igem.org/wiki/images/2/2a/T--Tuebingen--Gel2.png" | + | href="https://2019.igem.org/wiki/images/9/94/T--Tuebingen--Experimenta-Lab3.png" |
| | data-effect="mfp-zoom-out" | | data-effect="mfp-zoom-out" |
| | title="Some title"> | | title="Some title"> |
| − | <img src="https://2019.igem.org/wiki/images/2/2a/T--Tuebingen--Gel2.png" | + | <img src="https://2019.igem.org/wiki/images/9/94/T--Tuebingen--Experimenta-Lab3.png" |
| | class="figure-img img-fluid rounded" | | class="figure-img img-fluid rounded" |
| − | alt="A generic square placeholder image"></a> | + | alt="Placeholder"></a> |
| − | <figcaption style="color: #F4F0DE; font-size: small;">Figure 2. 07/11/19 Gelelctrophoresis of Cascade PCR. Gel loaded (from left to right): | + | </figure> |
| − | Ladder 1kb, Cascade (68°C), Cascade (64°C). It shows positive PCR of Cas 3 (68°C), expected at ~4.4 kb.</figcaption>
| + | |
| − | </figure>
| + | </div> |
| − | </div>
| + | <div class="col-12 col-md-6 "> |
| − | </div>
| + | <figure> |
| − | </div>
| + | |
| − | </div>
| + | |
| − | <div id="accordion2">
| + | |
| − | <div class="card">
| + | |
| − | <div class="card-header" id="headingOne">
| + | |
| − | <h5 class="mb-0">
| + | |
| − | <button class="btn btn-one btn-link collapsed" data-toggle="collapse" data-target="#collapseTwo" aria-expanded="false" aria-controls="collapseTwo">
| + | |
| − | Week 2 (15 Jul - 19 Jul)
| + | |
| − | </button> | + | |
| − | </h5>
| + | |
| − | </div>
| + | |
| − | <div id="collapseTwo" class="collapse" aria-labelledby="headingTwo" data-parent="#accordion2" >
| + | |
| − | <div class="card-body" style="background-color: #232323;">
| + | |
| − | <p style="color: #F4F0DE">
| + | |
| − | 15 Jakob Patrick Eva Luzi The biobricks were transformed again. Cascade and Cas 3 PCR products were
| + | |
| − | sent off for Sanger sequencing.
| + | |
| − | 16 Patrick, Eva Since there repeatedly weren’t any colonies for BBa_K608351 (K2), we transformed it
| + | |
| − | in provided NovaBlue™ competent cells. Overnight cultures of the other biobricks as well as an
| + | |
| − | overnight culture of E.Coli Nissle were inoculated. 17 Eva Patrick Katharina Glycerol
| + | |
| − | stocks of the overnight cultures were prepared. Competent E. Coli Nissle cells were produced and
| + | |
| − | the growth of E. Coli Nissle was analysed preliminarily. After Miniprep (Qiagen) of BBa_K516030 (RFP)
| + | |
| − | a PCR (Pfu Polymerase) was run. After digestion of the PCR products Cas 3 and Cascade as well as the
| + | |
| − | plasmid BBa_I13453 (K10), K10_Cascade and K10_Cas3 were ligated. 18 Eva Patrick The K10_Cascade and
| + | |
| − | K10_Cas3 ligations were digested (single) and analysed by gel electrophoresis, together with the RFP
| + | |
| − | PCR product (figure 3). The gel did not show a result for K10_Cascade and K10_Cas3. RFP (PCR Product)
| + | |
| − | and BBa_R0073 (Mnt Promotor) were digested and ligated. 19 Eva Patrick Digestion and Ligation of Cascade,
| + | |
| − | Cas3 and K10 were repeated with increased amounts of DNA. K10_Cascade, K10_Cas3 and BBa_R0073(Mnt)_RFP
| + | |
| − | were transformed.
| + | |
| − | </p>
| + | |
| − | <figure>
| + | |
| | <a style="font-size: small" | | <a style="font-size: small" |
| − | href="https://2019.igem.org/wiki/images/3/30/T--Tuebingen--Gel3.png" | + | href="https://2019.igem.org/wiki/images/6/69/T--Tuebingen--Experimenta-Lab5.png" |
| | data-effect="mfp-zoom-out" | | data-effect="mfp-zoom-out" |
| | title="Some title"> | | title="Some title"> |
| − | <img src="https://2019.igem.org/wiki/images/3/30/T--Tuebingen--Gel3.png" | + | <img src="https://2019.igem.org/wiki/images/6/69/T--Tuebingen--Experimenta-Lab5.png" |
| | class="figure-img img-fluid rounded" | | class="figure-img img-fluid rounded" |
| − | alt="A generic square placeholder"></a> | + | alt="Placeholder"></a> |
| − | <figcaption style="color: #F4F0DE; font-size: small;">Figure 3. 07/18/19 Gelelctrophoresis of | + | </figure> |
| − | RFP PCR, digests of K10_Cascade and K10_Cas3. Gel loaded (from left to right): Ladder 1 kb,
| + | </div> |
| − | RFP 1, RFP 2, RFP 3, K10_Cas3 (no heat inact.), K10_Cas3, K10_Cascade, Ladder 1 kb. It shows
| + | </div> |
| − | positive PCR of RFP, expected at ~890 bp. No result for digestions.</figcaption>
| + | <p> |
| − | </figure>
| + | Please check out the material we prepared for the course - a script covering all the theoretical |
| − | </div>
| + | background and a presentation: |
| − | </div>
| + | </p> |
| − | </div>
| + | <ul style="color:#F4F0DE;"> |
| − | </div> | + | <li><a href="https://2019.igem.org/wiki/images/6/6c/T--Tuebingen--Experimenta_Labkurs_Skript.pdf" onclick='window.open("https://2019.igem.org/wiki/images/6/6c/T--Tuebingen--Experimenta_Labkurs_Skript.pdf","_self");'>Experimenta Labkurs</a></li> |
| − | <div id="accordion3">
| + | <li><a href="https://2019.igem.org/wiki/images/1/10/T--Tuebingen--LabCoursePresentation.pdf" onclick='window.open("https://2019.igem.org/wiki/images/1/10/T--Tuebingen--LabCoursePresentation.pdf","_self");'>LabCourse Presentation</a>)</li> |
| − | <div class="card">
| + | </ul> |
| − | <div class="card-header" id="headingThree">
| + | <p> |
| − | <h5 class="mb-0">
| + | Please see the schedule below. |
| − | <button class="btn btn-one btn-link collapsed" data-toggle="collapse" data-target="#collapseThree" aria-expanded="false" aria-controls="collapseThree">
| + | </p> |
| − | Week 3 (22 Jul - 26 Jul)
| + | </div> |
| − | </button>
| + | </div> |
| − | </h5>
| + | |
| − | </div>
| + | <div class="row"> |
| − | <div id="collapseThree" class="collapse" aria-labelledby="headingThree" data-parent="#accordion3" >
| + | <div class="col-12"> |
| − | <div class="card-body" style="background-color: #232323;">
| + | <figure> |
| − | <p style="color: #F4F0DE">
| + | |
| − | 23 Eva Jakob Patrick Preparation of LB-Medium, LB-Agar and Chloramphenicol plates. Overnight cultures
| + | |
| − | for K10_Cascade, K10_Cas3 and BBa_R0073(Mnt)_RFP colonies were inoculated. After double digestion of
| + | |
| − | BBa_K584000 (AraC Promotor), BBa_K117008 (LsrR Promotor), the fragments pAraC and pLsrR were extracted
| + | |
| − | from the gel. 24 Eva Patrick After Miniprep of K10_Cascade, K10_Cas3 and BBa_R0073(Mnt)_RFP (Qiagen)
| + | |
| − | the plasmids were single digested. Gelelectrophoresis showed the empty backbone (figure 4).
| + | |
| − | The isolated fragments pAraC and pLsrR were ligated into the backbone psB1K3 after digestion.
| + | |
| − | psBIK3_pAraC and psBIK3_pLsrR were transformed. 25 Eva Overnight cultures for psBIK3_pAraC and psBIK3_pLsrR
| + | |
| − | colonies were inoculated. 26 Eva After Miniprep (Qiagen) of psBIK3_pAraC and psBIK3_pLsrR, the plasmids
| + | |
| − | were digested. Gel electrophoresis showed successful ligations (figure 5).
| + | |
| − | </p>
| + | |
| − | <figure>
| + | |
| | <a style="font-size: small" | | <a style="font-size: small" |
| − | href="https://2019.igem.org/wiki/images/1/13/T--Tuebingen--Gel4.png" | + | href="https://2019.igem.org/wiki/images/f/f1/T--Tuebingen--table_test.png" |
| | data-effect="mfp-zoom-out" | | data-effect="mfp-zoom-out" |
| | title="Some title"> | | title="Some title"> |
| − | <img src="https://2019.igem.org/wiki/images/1/13/T--Tuebingen--Gel4.png" | + | <img src="https://2019.igem.org/wiki/images/f/f1/T--Tuebingen--table_test.png" |
| | class="figure-img img-fluid rounded" | | class="figure-img img-fluid rounded" |
| − | alt="A generic square placeholder"></a> | + | alt="Placeholder"></a> |
| − | <figcaption style="color: #F4F0DE; font-size: small;">Figure 4. 07/24/19 Single digestions after | + | </figure> |
| − | Miniprep. Gel loaded (from left to right): Ladder 1kb, BBa_R0073(Mnt)_RFP 1.1, BBa_R0073(Mnt)_RFP
| + | </div> |
| − | 1.2, K10_Cas3 1.1-3.2, K10_Cascade 1.1-3.2. BBa_R0073(Mnt)_RFP expected at ~3 kb, K10_Cas 3 expected | + | </div> |
| − | at ~4.9 kb, K10_Cascade expected at ~6.6 kb.</figcaption>
| + | |
| − | </figure>
| + | <div class="row"> |
| − | <figure>
| + | <div class="col-12"> |
| | + | <p>Due to the interactive design of the course, we also discussed the experiments with the students |
| | + | and tried to summarize what we did on the blackboard:</p> |
| | + | |
| | + | <div class="row align-items-center"> |
| | + | <div class="col-12 col-md-6"> |
| | + | <figure> |
| | <a style="font-size: small" | | <a style="font-size: small" |
| − | href="https://2019.igem.org/wiki/images/b/b6/T--Tuebingen--Gel5.png" | + | href="https://2019.igem.org/wiki/images/b/be/T--Tuebingen--Experimenta-Lab1.png" |
| | data-effect="mfp-zoom-out" | | data-effect="mfp-zoom-out" |
| | title="Some title"> | | title="Some title"> |
| − | <img src="https://2019.igem.org/wiki/images/b/b6/T--Tuebingen--Gel5.png" | + | <img src="https://2019.igem.org/wiki/images/b/be/T--Tuebingen--Experimenta-Lab1.png" |
| | class="figure-img img-fluid rounded" | | class="figure-img img-fluid rounded" |
| − | alt="A generic square placeholder image"></a> | + | alt="Placeholder"></a> |
| − | <figcaption style="color: #F4F0DE; font-size: small;">Figure 5. 07/26/19 Single Digestions after | + | </figure> |
| − | Miniprep. Gel loaded (from left to right): psBIK3_pAraC 1-8, psB1K3_pLsrR 1-8, Ladder 1kb,
| + | |
| − | psB1K3 2.2-2.3, 3.1-3.2. Bands of ~3 kb suggest successful ligations.</figcaption>
| + | </div> |
| − | </figure>
| + | <div class="col-12 col-md-6 "> |
| − | </div>
| + | <figure> |
| − | </div>
| + | |
| − | </div>
| + | |
| − | </div>
| + | |
| − | <div id="accordion4">
| + | |
| − | <div class="card">
| + | |
| − | <div class="card-header" id="headingFour">
| + | |
| − | <h5 class="mb-0">
| + | |
| − | <button class="btn btn-one btn-link collapsed" data-toggle="collapse" data-target="#collapseFour" aria-expanded="false" aria-controls="collapseFour">
| + | |
| − | Week 4 (29 Jul - 02 Aug)
| + | |
| − | </button> | + | |
| − | </h5>
| + | |
| − | </div>
| + | |
| − | <div id="collapseFour" class="collapse" aria-labelledby="headingFour" data-parent="#accordion4" >
| + | |
| − | <div class="card-body" style="background-color: #232323;">
| + | |
| − | <p style="color: #F4F0DE">
| + | |
| − | 30 Patrick, Katharina The team moved into a new laboratory. The synthesized sequences
| + | |
| − | (K0, K3, K4, K6, K7, K15) arrived and were aliquoted. Chloramphenicol plates were prepared.
| + | |
| − | Cascade, Cas3, BBa_I13453 (K10), RFP and BBa_R0073 (Mnt Promotor) digestions and ligations were
| + | |
| − | repeated and transformed. 31 Patrick Jakob Overnight cultures for K10_Cas3 colonies were inoculated,
| + | |
| − | there was no growth of the other transformations. A PCR (Pfu polymerase) was run on the synthesized
| + | |
| − | constructs, it wasn’t successful. The Minipreps of psBIK3_pAraC and psBIK3_pLsrR were used for retransformation.
| + | |
| − | 01 Patrick The K10_Cas3 overnight culture was miniprepped. Overnight cultures for psBIK3_pAraC, psBIK3_pLsrR,
| + | |
| − | as well as K10_Cascade and BBa_R0073(Mnt)_RFP (some late colonies grew) were inoculated.
| + | |
| − | 02 Patrick Glycerol stocks for psBIK3_pAraC were prepared, there was no growth in psBIK3_pLsrR overnight
| + | |
| − | cultures. After Miniprep of rpsBIK3_pAraC, K10_Cascade and BBa_R0073(Mnt)_RFP the plasmids (also K10_Cas3)
| + | |
| − | were digested and a gel ectrophoresis was run. It showed the transformation of empty backbone in K10_Cascade
| + | |
| − | and BBa_R0073(Mnt)_RFP, successful ligation for psBIK3_pAraC and XY for K10_Cas3 (figure 6).
| + | |
| − | </p>
| + | |
| − | <figure>
| + | |
| | <a style="font-size: small" | | <a style="font-size: small" |
| − | href="https://2019.igem.org/wiki/images/8/82/T--Tuebingen--Gel6.png" | + | href="https://2019.igem.org/wiki/images/d/df/T--Tuebingen--Experimenta-Lab2.png" |
| | data-effect="mfp-zoom-out" | | data-effect="mfp-zoom-out" |
| | title="Some title"> | | title="Some title"> |
| − | <img src="https://2019.igem.org/wiki/images/8/82/T--Tuebingen--Gel6.png" | + | <img src="https://2019.igem.org/wiki/images/d/df/T--Tuebingen--Experimenta-Lab2.png" |
| | class="figure-img img-fluid rounded" | | class="figure-img img-fluid rounded" |
| − | alt="A generic square placeholder"></a> | + | alt="Placeholder"></a> |
| − | <figcaption style="color: #F4F0DE; font-size: small;">Figure 6. 08/02/19 Digestions after Miniprep. | + | </figure> |
| − | Gel loaded (from left to right): Ladder 1 kb, K10_Cas3 1-10, K10_Cascade 1-4, BBa_R0073(Mnt)_RFP 1-2,
| + | </div> |
| − | positive control, psBIK3_pAraC. Negative results for K10_Cas3, expected at ~4.9 kb, and K10_Cascade,
| + | </div> |
| − | expected at 6.6 kb. Successful ligation of psBIK3_pAraC, ~ 3 kb.</figcaption>
| + | </div> |
| − | </figure>
| + | </div> |
| − | </div>
| + | |
| − | </div>
| + | <div class="row"> |
| − | </div>
| + | <div class="col-12"> |
| − | </div> | + | <p>The students were also very interested in iGEM and our work. Hence, we not only introduced iGEM |
| − | <div id="accordion5">
| + | and our project to them, but also let them critically evaluate our project. They were also given |
| − | <div class="card">
| + | the chance to conduct PCRs related to our project and could therefore gain insights into the upcoming |
| − | <div class="card-header" id="headingFive">
| + | challenges when starting a new project or experiment where not everything has been optimised for |
| − | <h5 class="mb-0">
| + | easy functionality. Many students were interested in the iGEM competition itself and in the process |
| − | <button class="btn btn-one btn-link collapsed" data-toggle="collapse" data-target="#collapseFive" aria-expanded="false" aria-controls="collapseFive">
| + | of finding a project and making it feasible. Therefore, we also invested into giving valuable input |
| − | Week 5 (05 Aug - 09 Aug)
| + | and advice on starting a project and encouraged the students to participate in iGEM or similar projects |
| − | </button>
| + | themselves. Several questions and concerns relating the functionality of our probiotic under different |
| − | </h5>
| + | conditions, such as patients taking medications or antibiotics were also raised by the students. These |
| − | </div>
| + | fruitful discussions, as well as our exchange with several experts, contributed to the final design |
| − | <div id="collapseFive" class="collapse" aria-labelledby="headingThree" data-parent="#accordion5" >
| + | of our RNA-Seq experiment providing valuable input and different aspects to be considered |
| − | <div class="card-body" style="background-color: #232323;">
| + | [<a href="https://2019.igem.org/Team:Tuebingen/Experts" onclick='window.open("https://2019.igem.org/Team:Tuebingen/Experts","_self");'>Experts</a>, <a href="https://2019.igem.org/Team:Tuebingen/Nissle" onclick='window.open("https://2019.igem.org/Team:Tuebingen/Nissle","_self");'>Nissle</a>].</p> |
| − | <p style="color: #F4F0DE">
| + | |
| − | 05 Patrick Eva Antonia PCR (Q5 polymerase) and PCR (Paq polymerase) was run on the synthesized constructs,
| + | <p> |
| − | they weren’t successful. Cascade and Cas3 were successfully amplified via PCR(Q5). 06 Eva Patrick Antonia
| + | Due to the high interest of the students and the positive feedback from the Experimenta itself, |
| − | A gradient PCR (Q5) was run on the constructs. It was discovered that the currently used agarose was falsely
| + | we discussed the possibility of creating an iGEM high school team in Heilbronn in 2020. |
| − | prepared with water instead of TAE-Buffer and was discarded. Cascade, Cas3 and BBa_I13453 (K10) were
| + | The Experimenta is highly interested in participating in iGEM 2020 with motivated students and |
| − | digested and ligated. 07 Patrick Eva Luzi Antonia A new PCR (Q5) was run on the synthesized constructs,
| + | will provide the required laboratories, supervision and funding. Our team will still be serving |
| − | gelectrophoresis suggested positive apmlification (figure 7). By amplification with accordingly designed primer,
| + | as advisors for the planned iGEM high school team next year. Furthermore, a new student lab course |
| − | K0 has been added a TetR promotor and is now K1. Due to a technical malfunction in the building further
| + | on CRISPR/Cas will be introduced at the Experimenta on the grounds of the theoretical background, |
| − | work was delayed.
| + | input and inspiration iGEM Tuebingen conveyed. |
| − | </p>
| + | </p> |
| − | <figure>
| + | </div> |
| | + | </div> |
| | + | |
| | + | <h3 style="font-family:'Righteous'; color:#2ecc71;">Panel Discussion - „Biohacking – genetic experiments done at home“</h3> |
| | + | <div class="row"> |
| | + | <div class="col-12"> |
| | + | <p>In addition to the lab course, we also decided to host a panel discussion at the Experimenta. |
| | + | At the beginning of the <a href="https://2019.igem.org/wiki/images/2/24/T--Tuebingen--panelDiscussion.pdf" onclick='window.open("https://2019.igem.org/wiki/images/2/24/T--Tuebingen--panelDiscussion.pdf","_self");'>panel discussion</a>, we gave a talk about on the CRISPR / Cas method itself. |
| | + | We also included an overview of our ongoing project and the iGEM competition in general.</p> |
| | + | |
| | + | <p> |
| | + | In collaboration with Dr. Robert Friedrich, we designed a discussion around the status of current |
| | + | research on CRISPR / Cas, the effectiveness of the method and its limitations, the prospects of |
| | + | future CRISPR / Cas developments, the availability and legality of so-called Do-It-Yourself CRISPR / |
| | + | Cas Kits and the ethical implications on society. The panel consisted out of Prof. Dr. Christoph |
| | + | Thyssen (<a href="www.bio.uni-kl.de/organisation/mitarbeiter/s-u/thyssen-christoph/" onclick='window.open("www.bio.uni-kl.de/organisation/mitarbeiter/s-u/thyssen-christoph/","_self");'>Christoph Thyssen</a>), an expert for subject |
| | + | didactics in biology and two of our team members - Marietheres Evers and Famke Baeuerle. |
| | + | </p> |
| | + | |
| | + | <div class="row align-items-center"> |
| | + | <div class="col-12 col-md-6"> |
| | + | <figure> |
| | <a style="font-size: small" | | <a style="font-size: small" |
| − | href="https://2019.igem.org/wiki/images/4/46/T--Tuebingen--Gel7.png" | + | href="https://2019.igem.org/wiki/images/5/54/T--Tuebingen--Biohacking_05.jpg" |
| | data-effect="mfp-zoom-out" | | data-effect="mfp-zoom-out" |
| | title="Some title"> | | title="Some title"> |
| − | <img src="https://2019.igem.org/wiki/images/4/46/T--Tuebingen--Gel7.png" | + | <img src="https://2019.igem.org/wiki/images/5/54/T--Tuebingen--Biohacking_05.jpg" |
| | class="figure-img img-fluid rounded" | | class="figure-img img-fluid rounded" |
| − | alt="A generic square placeholder"></a> | + | alt="Placeholder"></a> |
| − | <figcaption style="color: #F4F0DE; font-size: small;">Figure 7. 08/08/19 PCR of synthesized constructs. | + | </figure> |
| − | Gel loaded (from left to right): Ladder 1kb, K1 (expected at 660bp), K3 (expected at 1 kb),
| + | |
| − | K4 (expected at 1 kb), K6 (expected at 1.5 kb), K7 (expected at 700 bp), K15 (expected at 1 kb).</figcaption>
| + | </div> |
| − | </figure>
| + | <div class="col-12 col-md-6 "> |
| − | </div>
| + | <figure> |
| − | </div>
| + | |
| − | </div>
| + | |
| − | </div>
| + | |
| − | <div id="accordion6">
| + | |
| − | <div class="card">
| + | |
| − | <div class="card-header" id="headingSix">
| + | |
| − | <h5 class="mb-0">
| + | |
| − | <button class="btn btn-one btn-link collapsed" data-toggle="collapse" data-target="#collapseSix" aria-expanded="false" aria-controls="collapseSix">
| + | |
| − | Week 6 (12 Aug - 16 Aug)
| + | |
| − | </button> | + | |
| − | </h5>
| + | |
| − | </div>
| + | |
| − | <div id="collapseSix" class="collapse" aria-labelledby="headingSix" data-parent="#accordion6" >
| + | |
| − | <div class="card-body" style="background-color: #232323;">
| + | |
| − | <p style="color: #F4F0DE">
| + | |
| − | 12 Patrick Katharina After respective digestion and DNA clean up, psBC13_K1, psBC13_K3, psBC13_K6, K5_K4,
| + | |
| − | K10_Cas3, K10_Cascade and BBa_R0073(Mnt)_RFP were ligated. 13 Patrick Eva Antonia Marie Yesterday’s ligations
| + | |
| − | were transformed. 14 Luzie Marie Eva A colony PCR (Taq polymerase) was run on psBC13_K1,
| + | |
| − | but the gelelectrophoresis was negative (figure 8). Overnight cultures of psBC13_K1, psBC13_K3, psBC13_K6,
| + | |
| − | K5_K4, K10_Cas3 and K10_Cascade colonies were inoculated. 15 Marie Patrick Eva After Miniprep the plasmids
| + | |
| − | were double digested. Gel electrophoresis suggested successful ligations of psBC13_K6, K10_Cas3, K10_Cascade
| + | |
| − | and BBa_R0073(Mnt)_RFP (figure 9). 16 Patrick Miniprep of psBC13_K1 was digested but gelectrophoresis was negative.
| + | |
| − | Cascade and Cas3 were amplified by PCR (Q5 polymerase). The synthesized construct K2 arrived, was diluted and
| + | |
| − | amplified via PCR (Q5 polymerase).
| + | |
| − | </p>
| + | |
| − | <figure>
| + | |
| | <a style="font-size: small" | | <a style="font-size: small" |
| − | href="https://2019.igem.org/wiki/images/2/25/T--Tuebingen--Gel8.png" | + | href="https://2019.igem.org/wiki/images/4/4d/T--Tuebingen--Biohacking_10.jpg" |
| | data-effect="mfp-zoom-out" | | data-effect="mfp-zoom-out" |
| | title="Some title"> | | title="Some title"> |
| − | <img src="https://2019.igem.org/wiki/images/2/25/T--Tuebingen--Gel8.png" | + | <img src="https://2019.igem.org/wiki/images/4/4d/T--Tuebingen--Biohacking_10.jpg" |
| | class="figure-img img-fluid rounded" | | class="figure-img img-fluid rounded" |
| − | alt="A generic square placeholder"></a> | + | alt="Placeholder"></a> |
| − | <figcaption style="color: #F4F0DE; font-size: small;">Figure 8. 08/14/19 Colony PCR of psB1C3_K1 | + | </figure> |
| − | transformation. Gel loaded (from left to right): Ladder 1kb, psB1C3_K0 1-10.</figcaption>
| + | </div> |
| − | </figure>
| + | </div> |
| − | <figure>
| + | <p> |
| − | <a style="font-size: small"
| + | A lot of questions were asked during the <a href="https://2019.igem.org/wiki/images/1/13/T--Tuebingen--Panel_discussion_allg.pdf" onclick='window.open("https://2019.igem.org/wiki/images/1/13/T--Tuebingen--Panel_discussion_allg.pdf","_self");'>panel discussion</a> and the presentation. We discussed |
| − | href="https://2019.igem.org/wiki/images/c/cc/T--Tuebingen--Gel9.png"
| + | the ethical implications of the CRISPR / Cas Method with the audience. Surprisingly, |
| − | data-effect="mfp-zoom-out"
| + | many participants were eager to talk about scenarios they have heard about. |
| − | title="Some title">
| + | </p> |
| − | <img src="https://2019.igem.org/wiki/images/c/cc/T--Tuebingen--Gel9.png"
| + | </div> |
| − | class="figure-img img-fluid rounded"
| + | </div> |
| − | alt="A generic square placeholder image"></a>
| + | |
| − | <figcaption style="color: #F4F0DE; font-size: small;">Figure 9. 08/15/19 Digestions after Miniprep. | + | |
| − | Gel loaded (from left to right): Ladder 1kb, psB1C3_K6 1, empty, psB1C3_K6 2, psB1C3_K3 1-2, K10_Cas3
| + | |
| − | 1-2, K10_Cascade 1-2, BBa_R0073(Mnt)_RFP 1-4. K6 expected at ~2 kb and 1.5 kb, K3 expected at
| + | |
| − | ~2 kb and 1 kb, Cas3 expected at ~2.4 kb, ~2 kb and ~0.3 kb, Cascade expected at ~5.1 kb, ~1,2 kb
| + | |
| − | and ~0.3 kb, RFP expected at ~2 kb and ~1 kb. </figcaption>
| + | |
| − | </figure>
| + | |
| − | </div>
| + | |
| − | </div>
| + | |
| − | </div>
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| − | </div>
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| − | <div id="accordion7">
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| − | <div class="card">
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| − | <div class="card-header" id="headingSeven">
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| − | <h5 class="mb-0">
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| − | <button class="btn btn-one btn-link collapsed" data-toggle="collapse" data-target="#collapseSeven" aria-expanded="false" aria-controls="collapseSeven">
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| − | Week 7 (19 Aug - 23 Aug)
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| − | </button>
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| − | </h5>
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| − | </div>
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| − | <div id="collapseSeven" class="collapse" aria-labelledby="headingSeven" data-parent="#accordion7" >
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| − | <div class="card-body" style="background-color: #232323;">
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| − | <p style="color: #F4F0DE">
| + | |
| − | 19 Patrick Jakob Eva After respective digestion and DNA clean up, psB1C3_K1, psB1C3_K3, psB1C3_K7 and
| + | |
| − | K5_K4 were ligated. The BBa_R0073(Mnt)_RFP plasmid was digested to isolate Mnt_RFP by gel extraction
| + | |
| − | (figure 10). The backbone pSB1K3 was digested with respective enzymes and pSB1K3_Mnt_RFP was ligated.
| + | |
| − | K1, K2, K3, K4, K7 and K15 were amplified via PCR (Q5 polymerase) with a new programm. 20 Eva Patrick
| + | |
| − | Last weeks psBC13_K6, K10_Cas3 and K10_Cascade were sent off to Sanger sequencing. Yesterday’s ligations
| + | |
| − | were transformed as well as K10_Cascade retransformed. 21 Marie Zoe Patrick A new batch of competent cells
| + | |
| − | was prepared and overnight cultures of psB1C3_K1, psB1C3_K7, K5_K4 and K10_Cascade inoculated. 22 Patrick
| + | |
| − | Eva Sanger Sequencing was positive on K10_Cascade. Chloramphenicol and Kanamycin plates were prepared.
| + | |
| − | After Miniprep of psB1C3_K1, psB1C3_K7, K5_K4 and K10_Cascade the plasmids were digested and run on a gel.
| + | |
| − | Digestions of psB1C3_K1 and K5_K4 were negative. The new competent cells were transformed with psB1C3_K3,
| + | |
| − | psB1K3_Mnt_RFP and iGEM competent cell test plasmids. 23 Marie Patrick The constructs K1, K2, K3, K4, K7
| + | |
| − | and K15 were amplified by PCR (Q5 polymerase). Double digestion of psB1C3_K7 and K10_Cascade was repeated.
| + | |
| − | The gel electrophoresis suggested XY (figure 11).
| + | |
| − | </p>
| + | |
| − | <figure>
| + | |
| − | <a style="font-size: small"
| + | |
| − | href="https://2019.igem.org/wiki/images/7/77/T--Tuebingen--Gel10.png"
| + | |
| − | data-effect="mfp-zoom-out"
| + | |
| − | title="Some title">
| + | |
| − | <img src="https://2019.igem.org/wiki/images/7/77/T--Tuebingen--Gel10.png"
| + | |
| − | class="figure-img img-fluid rounded"
| + | |
| − | alt="A generic square placeholder"></a>
| + | |
| − | <figcaption style="color: #F4F0DE; font-size: small;">Figure 10. 08/19/19 Digestion of BBa_R0073(Mnt)_RFP
| + | |
| − | for gel extraction. Gel loaded (from left to right): Ladder 1 kb, BBa_R0073(Mnt)_RFP. | + | |
| − | Desired fragment at ~0.5 kb.</figcaption> | + | |
| − | </figure>
| + | |
| − | <figure>
| + | |
| − | <a style="font-size: small"
| + | |
| − | href="https://2019.igem.org/wiki/images/9/92/T--Tuebingen--Gel11.png"
| + | |
| − | data-effect="mfp-zoom-out"
| + | |
| − | title="Some title">
| + | |
| − | <img src="https://2019.igem.org/wiki/images/9/92/T--Tuebingen--Gel11.png"
| + | |
| − | class="figure-img img-fluid rounded"
| + | |
| − | alt="A generic square placeholder image"></a>
| + | |
| − | <figcaption style="color: #F4F0DE; font-size: small;">Figure 11. Digestion after Miniprep. | + | |
| − | Gel loaded (from left to right): Ladder 1 kb, psB1C3_K7 1-2, K10_Cascade 1-5, K10_Cas3.</figcaption>
| + | |
| − | </figure>
| + | |
| − | </div>
| + | |
| − | </div>
| + | |
| − | </div>
| + | |
| − | </div> | + | |
| − | </div> | + | |
| | | | |
| | + | <h3 style="font-family:'Righteous'; color:#2ecc71;">Outlook - Advisors of the Experimenta iGEM team 2020 & New student lab course</h3> |
| | + | <div class="row"> |
| | + | <div class="col-12"> |
| | + | <p>During the collaboration with the Experimenta Heilbronn, we sat together with Dr. Robert Friedrich |
| | + | and Norbert Hommrichhausen to discuss the foundation of an iGEM high school team at the Experimenta |
| | + | Heilbronn. The Experimenta is highly interested in starting a team in Heilbronn and the motivated |
| | + | and interested students encouraged us to team up and further pursue this plan. Plans for an iGEM |
| | + | high school team 2020 were made, in which the Experimenta Heilbronn will be the main partner |
| | + | providing lab space, supervision and funding. The role of the iGEM Team Tuebingen is that of |
| | + | experienced advisors, who can assist in questions concerning the iGEM competition itself, the |
| | + | workflow of an iGEM year, as well as deliver expert knowledge and guidance in the laboratory or |
| | + | in questions concerning bioinformatics, design and human practices. Both the Experimenta and the |
| | + | iGEM Team Tuebingen aim at establishing a long-term collaboration in the scope of iGEM.</p> |
| | | | |
| − | | + | <p> |
| − | </div>
| + | Next to the foundation of a high school iGEM team, the Experimenta will also introduce new lab |
| − | | + | courses for school classes, one of them being a CRISPR/Cas course. The theoretical background, |
| − | <div id="glpreferences" class="row">
| + | first steps and foundation of this course arose from the collaboration with iGEM Tuebingen this year. |
| − | <h3 style="font-family:'Righteous'; color: #2ecc71;">References</h3>
| + | A more detailed outline and implementation of the course will be part of another collaboration |
| − | <div class="col-12" style="font-size: x-small; overflow-wrap: break-word;">
| + | between the Experimenta and iGEM Tuebingen. |
| − | <ol> | + | </p> |
| − | <li style="color: #F4F0DE"><a href=https://doi.org/10.1093/femsle/fnw212> Ulrich Sonnenborn, Escherichia coli strain Nissle 1917—from bench to bedside and back: history of a special Escherichia coli strain with probiotic properties, FEMS Microbiology Letters, Volume 363, Issue 19, October 2016, fnw212,</a></li> | + | |
| − | <li id="ref_2" style="color: #F4F0DE">Sana Ben Othman and Tomio Yabe. Use of Hydrogen Peroxide and Peroxl Radicals to induce Oxidative Stress in Neuronal Cells. Reviews in Agricultural Science, 3:40-45, 2015. Doi: 10.7831/ras.3.40</li> | + | |
| − | <li id="ref_3" style="color: #F4F0DE;">P. Capela, T.K.C. Hay, N.P. Shah. Effect of cryoprotectants, prebiotics and microencapsulation on survival of probiotic organisms in yoghurt and freeze-dried yoghurt. Food Research International. Volume 39, Issue 2, 2006, Pages 203-211, ISSN 0963-9969, https://doi.org/10.1016/j.foodres.2005.07.007.</li>
| + | |
| − | <li id="ref_4" style="color: #F4F0DE;">Govender M, Choonara YE, Kumar P, du Toit LC, van Vuuren S, Pillay V. A review of the advancements in probiotic delivery: Conventional vs. non-conventional formulations for intestinal flora supplementation. AAPS PharmSciTech. 2014;15(1):29–43. doi:10.1208/s12249-013-0027-1</li>
| + | |
| − | <li id="ref_5" style="color: #F4F0DE;"><a href="https://opsdiagnostics.com/applications/lyophilization/ecoli_lyophilization_stability.html">https://opsdiagnostics.com/applications/lyophilization/ecoli_lyophilization_stability.html</a> </li> | + | |
| − | <li id="ref_6" style="color: #F4F0DE;"><a href="https://doi.org/10.1007/s00125-008-1053-5"></a>Bailey, C. J, Wilcock, C, Scarpello, J. H. B. Metformin and the intestine. Diabetologia. 2008. Volume 51, Issue 8, Pages 1552, ISSN 1432-0428, Doi: 10.1007/s00125-008-1053-5,</li>
| + | |
| − | <li id="ref_7" style="color: #F4F0DE;">High Accumulation of Metformin in Colonic Tissue of Subjects With Diabetes or the Metabolic Syndrome. Paleari, LauraBurhenne, JürgenFoersch, SebastianParodi, AndreaGnant, MichaelScherer, DominiqueUlrich, Cornelia M.Stabuc, BorutPuntoni, MatteoCoccia, GianniPetrera, MarilenaHaefeli, Walter-EmilDeCensi, Andrea et al. Gastroenterology, Volume 154, Issue 5, 1543 - 1545</li>
| + | |
| − | <li id="ref_8" style="color: #F4F0DE;">Martinez-Augustin O, Sanchez de Medina F. Intestinal bile acid physiology and pathophysiology. World J Gastroenterol. 2008;14(37):5630–5640. doi:10.3748/wjg.14.5630</li> | + | |
| − | <li id="ref_9" style="color: #F4F0DE;">Postprandial concentrations of free and conjugated bile acids down the length of the normal human small intestine. T. C. NORTHFIELD AND I. McCOLL. Gut, 1973, 14, 513-518</li>
| + | |
| − | <li id="ref_10" style="color: #F4F0DE;">Belizário JE, Napolitano M. Human microbiomes and their roles in dysbiosis, common diseases, and novel therapeutic approaches. Front Microbiol. 2015;6:1050. Published 2015 Oct 6. doi:10.3389/fmicb.2015.01050</li> | + | |
| − | <li id="ref_11" style="color: #F4F0DE;">Lloyd-Price J, Abu-Ali G, Huttenhower C. The healthy human microbiome. Genome Med. 2016;8(1):51. Published 2016 Apr 27. doi:10.1186/s13073-016-0307-y</li>
| + | |
| − | </ol> | + | |
| | </div> | | </div> |
| | </div> | | </div> |
| | + | </div> |
| | </main> | | </main> |
| | + | </div> |
| | </div> | | </div> |
| | | | |