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| | <ul class="main-menue-ul"> | | <ul class="main-menue-ul"> |
| | <a href="https://2014.igem.org/Team:Berlin" class="main-menue-links"><li>Home</li></a> | | <a href="https://2014.igem.org/Team:Berlin" class="main-menue-links"><li>Home</li></a> |
| - | <a href="https://2014.igem.org/Team:Berlin/Project" class="main-menue-links"><li>Project</li></a> | + | <a href="https://2014.igem.org/Team:Berlin/Project" class="main-menue-links"><li class="active">Project</li></a> |
| | <a href="https://2014.igem.org/Team:Berlin/Team" class="main-menue-links"><li>Team</li></a> | | <a href="https://2014.igem.org/Team:Berlin/Team" class="main-menue-links"><li>Team</li></a> |
| - | <a href="https://2014.igem.org/Team:Berlin/Safety" class="main-menue-links"><li>Safety</li></a> | + | <a href="https://2014.igem.org/Team:Berlin/Safety" class="main-menue-links"><li>Safety</li></a> |
| | <a href="https://2014.igem.org/Team:Berlin/Workshop" class="main-menue-links"><li>Workshop</li></a> | | <a href="https://2014.igem.org/Team:Berlin/Workshop" class="main-menue-links"><li>Workshop</li></a> |
| - | <a href="https://2014.igem.org/Team:Berlin/Blog"><li>Blog</li></a> | + | <a href="https://2014.igem.org/Team:Berlin/Blog" class="main-menue-links"><li>Blog</li></a> |
| | <a href="https://igem.org/Main_Page" class="igem-link"><li><img class="igem-logo" src="https://static.igem.org/mediawiki/2014/d/d8/Team_Berlin_igem_logo.png"></li></a> | | <a href="https://igem.org/Main_Page" class="igem-link"><li><img class="igem-logo" src="https://static.igem.org/mediawiki/2014/d/d8/Team_Berlin_igem_logo.png"></li></a> |
| | </ul> | | </ul> |
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| | <div class="row featurette" id="main-featurette-row"> | | <div class="row featurette" id="main-featurette-row"> |
| | | | |
| - | <div class="start-text-container"> | + | <div class="start-text-container-project"> |
| | <h2><img src="https://static.igem.org/mediawiki/2014/f/ff/Team_Berlin_igem_questionmark.png" alt="" class="teaser-icons hidden-xs" />Explore our Project:</h2> | | <h2><img src="https://static.igem.org/mediawiki/2014/f/ff/Team_Berlin_igem_questionmark.png" alt="" class="teaser-icons hidden-xs" />Explore our Project:</h2> |
| | <div class="teaser-text-sub"> | | <div class="teaser-text-sub"> |
| - | <div class="col-xs-12">
| |
| - | <a name="top"> </a>
| |
| - | <a href="#description" class="sub-link-project"> 1. What is it all about?</a><br/>
| |
| - | <a href="#animation" class="sub-link-project">2. Animation</a><br/>
| |
| - | <a href="#detail-description" class="sub-link-project">3. Detailed description of the different strategies</a><br/>
| |
| - | <a href="#flow-chart" class="sub-link-project">4. Flow Chart with the strategies</a><br/>
| |
| - | <a href="#results" class="sub-link-project">5) Our Results</a><br/>
| |
| - | <a href="#lab-summary" class="sub-link-project">6. Lab-summary</a><br/>
| |
| - | <a href="#notebook" class="sub-link-project">7. Notebook</a><br/>
| |
| - | <br/>
| |
| - | </div>
| |
| | </div> | | </div> |
| | <!--<a href="#" class="more-informations-white">More informations</a>--> | | <!--<a href="#" class="more-informations-white">More informations</a>--> |
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| | <!--- ////////////////// DESCRIPTION////////////////// --> | | <!--- ////////////////// DESCRIPTION////////////////// --> |
| | <div class="container"> | | <div class="container"> |
| - | <div class="row hidden-xs"> | + | |
| - | | + | <div class="col-xs- 3 submenue-project" style="text-align:left;"> |
| - | <div class="project-entry-tags">
| + | <a href="https://2014.igem.org/Team:Berlin/Project" class="sub-link-project"> 1. What is it all about?</a><br/><br/> |
| - | <ul>
| + | <a href="https://2014.igem.org/Team:Berlin/Project/Activities" class="sub-link-project"> 2. Project-related Activities</a><br/><br/> |
| - | < li><a href=" #top"> GO TO TOP</a></ li> | + | <a href="https://2014.igem.org/Team:Berlin/Project/Detailed-Description" class="sub-link-project">3. Detailed Description</a><br/><br/> |
| - | </ul>
| + | <a href="https://2014.igem.org/Team:Berlin/Project/Results" class="sub-link-project">4. Our Results</a><br/><br/> |
| - | </div>
| + | <a href=" https://2014.igem.org/Team:Berlin/Project/Summary" class="sub-link-project"> 5. Lab Summary</a>< br/><br/> |
| - | </div>
| + | <a href="https://2014.igem.org/Team:Berlin/Project/Journal" class="sub-link-project">6. Lab Journal</a><br/><br/> |
| - | </div>
| + | <a href="https://2014.igem.org/Team:Berlin/Project/Property" class="sub-link-project">7. Intellectual Property</a><br/><br/> |
| | + | <br/> |
| | + | </div> |
| | | | |
| - | <div class="row">
| + | <div class="col-xs-11 col-sm-9 blog-text" style="margin-bottom:40px;"> |
| - | <div class="col-xs-12"> | + | |
| | <div class="project-number">1</div><div class="project-headline-float"><h2 class="green-text project-headline"> What is it all about?</h2></div> | | <div class="project-number">1</div><div class="project-headline-float"><h2 class="green-text project-headline"> What is it all about?</h2></div> |
| - | </div>
| + | |
| - | </div>
| + | |
| - |
| + | |
| - | <div class="row">
| + | |
| - | <!--<div class="col-sm-4 hidden-xs">
| + | |
| - | <img src="img/blog/Team_Berlin_bacteria.png" class="img-responsive" style="margin: 0 auto;"/>
| + | |
| - | </div>-->
| + | |
| - | <div class="col-xs-12 col-sm-12 blog-text" style="margin-bottom:40px;">
| + | |
| | <p> | | <p> |
| | <a name="description"> </a> | | <a name="description"> </a> |
| - | As previous iGEM teams have shown, synthesizing fully functional magnetosomes in E. coli is highly difficult as more than 60 highly regulated genes are involved. As a more feasible alternative, we simply want to synthesize magnetic nanoparticles in E. coli in order to attract cells with strong magnetic fields.<br/>
| + | |
| - | Therefore we want to use different strategies including manipulation of the iron homeostasis of E. coli, expression of different metal binding proteins such as ferritins and metallothioneins as well as a high-throughput growth medium optimization.
| + | <strong>iGEM Berlin 2014: A remote control for E. coli</strong><br/> |
| - | <br/><br/>
| + | <p> |
| - | Furthermore, we will work with other metal binding proteins such as metallothioneins and phytochelatin synthases in order to achieve nanoparticle synthesis.
| + | As the first iGEM team from Berlin, we decided to construct a simple BioBrick that enables synthetic biologists to remotely control the movement of E. coli. A seemingly simple and non-invasive mechanism for this remote control is the use of magnetic fields. By altering the iron homeostasis of E. coli, we want to increase the total iron level of the cytosol. By sequestering iron in a ferritin protein, iron crystals are formed and the cell is detoxified. We also worked with other metal binding proteins such as metallothioneins and phytochelatin synthases in order to create various metal nanoparticles as an alternative strategy. Furthermore, we searched for the optimal conditions which yielded the most efficient formation of magnetic nanoparticles in E. coli. Once we have discovered the best way to magnetize E. coli bacteria, we will build and characterize suitable BioBricks that can be used to remote control the cellular movement of E. coli.</p><br/> |
| - | Once we have discovered the best way to magnetize E. coli bacteria, we will build and characterize suitable BioBricks that can be used by any research lab or iGEM team in the world in order to remote control the cellular movement.</p>
| + | The following animation visualizes the concept of using ferritin iron storage proteins as a magnetism mediating module.<br/> |
| - | </div>
| + | <br/> |
| - | </div>
| + | <iframe src="//player.vimeo.com/video/109249906?byline=0&portrait=0&color=0ecd28" width="700" height="394" frameborder="0" webkitallowfullscreen mozallowfullscreen allowfullscreen></iframe> |
| - | </div> | + | <br/> |
| - | <!--- ////////////////// ANIMATION ////////////////// --> | + | <br/> |
| - | <div class="container">
| + | This animation was developed by Florian Renner who is an science interested and very talented graphic designer based in Munich. We thank Florian for his amazing job he did voluntarily for iGEM Berlin helping us to overcome the limitations of our budget. |
| - | <div class="row hidden-xs">
| + | <br/><br/> |
| - | <div class="col-xs-12">
| + | |
| - | <div class="project-entry-tags">
| + | |
| - | <ul>
| + | |
| - | <li><a href="#top">GO TO TOP</a></li>
| + | |
| - | </ul>
| + | |
| - | </div>
| + | |
| - | </div>
| + | |
| - | </div>
| + | |
| - |
| + | |
| - | <div class="row">
| + | |
| - | <div class="col-xs-12">
| + | |
| - | <div class="project-number">2</div><div class="project-headline-float"><h2 class="green-text project-headline"> Visualisation</h2></div>
| + | |
| - | </div>
| + | |
| - | </div>
| + | |
| - |
| + | |
| - | <div class="row">
| + | |
| - | <!--<div class="col-sm-4 hidden-xs">
| + | |
| - | <img src="img/blog/Team_Berlin_bacteria.png" class="img-responsive" style="margin: 0 auto;"/> | + | |
| - | </div>-->
| + | |
| - | <div class="col-xs-12 col-sm-12 blog-text" style="margin-bottom:40px;">
| + | |
| - | <p>
| + | |
| - | <a name="animation"> </a>
| + | |
| - | Animation, jo!
| + | |
| - | </div>
| + | |
| - | </div>
| + | |
| - | </div>
| + | |
| | | | |
| - | <!--- ////////////////// DETAILED DESCRIPTION ////////////////// -->
| + | </div> |
| - | <div class="container">
| + | |
| - | <div class="row hidden-xs">
| + | |
| - | <div class="col-xs-12">
| + | |
| - | <div class="project-entry-tags">
| + | |
| - | <ul>
| + | |
| - | <li><a href="#top">GO TO TOP</a></li>
| + | |
| - | </ul>
| + | |
| - | </div>
| + | |
| - | </div>
| + | |
| - | </div>
| + | |
| - |
| + | |
| - | <div class="row">
| + | |
| - | <div class="col-xs-12">
| + | |
| - | <div class="project-number">3</div><div class="project-headline-float"><h2 class="green-text project-headline"> Detailes Description</h2></div>
| + | |
| - | </div>
| + | |
| - | </div>
| + | |
| - |
| + | |
| - | <div class="row">
| + | |
| - | <!--<div class="col-sm-4 hidden-xs">
| + | |
| - | <img src="img/blog/Team_Berlin_bacteria.png" class="img-responsive" style="margin: 0 auto;"/>
| + | |
| - | </div>-->
| + | |
| - | <div class="col-xs-12 col-sm-12 blog-text" style="margin-bottom:40px;">
| + | |
| - | <p>
| + | |
| - | <a name="detail-description"> </a>
| + | |
| - | MORE DETAILS, jo!
| + | |
| - | </div>
| + | |
| | </div> | | </div> |
| | </div> | | </div> |
| - | | + | |
| - | <!--- ////////////////// FLOW CHART ////////////////// -->
| + | </section> |
| - | <div class="container">
| + | |
| - | <div class="row hidden-xs">
| + | |
| - | <div class="col-xs-12">
| + | |
| - | <div class="project-entry-tags">
| + | |
| - | <ul>
| + | |
| - | <li><a href="#top">GO TO TOP</a></li>
| + | |
| - | </ul>
| + | |
| - | </div>
| + | |
| - | </div>
| + | |
| - | </div>
| + | |
| - |
| + | |
| - | <div class="row">
| + | |
| - | <div class="col-xs-12">
| + | |
| - | <div class="project-number">4</div><div class="project-headline-float"><h2 class="green-text project-headline"> Flow Chart</h2></div>
| + | |
| - | </div>
| + | |
| - | </div>
| + | |
| - |
| + | |
| - | <div class="row">
| + | |
| - | <!--<div class="col-sm-4 hidden-xs">
| + | |
| - | <img src="img/blog/Team_Berlin_bacteria.png" class="img-responsive" style="margin: 0 auto;"/>
| + | |
| - | </div>-->
| + | |
| - | <div class="col-xs-12 col-sm-12 blog-text" style="margin-bottom:40px;">
| + | |
| - | <p>
| + | |
| - | <a name="flow-chart"> </a>
| + | |
| - | <img src="https://static.igem.org/mediawiki/2014/e/ee/Team_berlin_flow-chart-01.png" alt="" />
| + | |
| - | </div>
| + | |
| - | </div>
| + | |
| - | </div>
| + | |
| - | | + | |
| - | <!--- ////////////////// Results ////////////////// -->
| + | |
| - | <div class="container">
| + | |
| - | <div class="row hidden-xs">
| + | |
| - | <div class="col-xs-12">
| + | |
| - | <div class="project-entry-tags">
| + | |
| - | <ul>
| + | |
| - | <li><a href="#top">GO TO TOP</a></li>
| + | |
| - | </ul>
| + | |
| - | </div>
| + | |
| - | </div>
| + | |
| - | </div>
| + | |
| - |
| + | |
| - | <div class="row">
| + | |
| - | <div class="col-xs-12">
| + | |
| - | <div class="project-number">5</div><div class="project-headline-float"><h2 class="green-text project-headline"> The Results</h2></div>
| + | |
| - | </div>
| + | |
| - | </div>
| + | |
| - |
| + | |
| - | <div class="row">
| + | |
| - | <!--<div class="col-sm-4 hidden-xs">
| + | |
| - | <img src="img/blog/Team_Berlin_bacteria.png" class="img-responsive" style="margin: 0 auto;"/>
| + | |
| - | </div>-->
| + | |
| - | <div class="col-xs-12 col-sm-12 blog-text" style="margin-bottom:40px;">
| + | |
| - | <p>
| + | |
| - | <a name="results"> </a>
| + | |
| - | Results, jo!
| + | |
| - | </div>
| + | |
| - | </div>
| + | |
| - | </div>
| + | |
| - | | + | |
| - | <!--- ////////////////// Lab Diary ////////////////// -->
| + | |
| - | <div class="container">
| + | |
| - | <div class="row hidden-xs">
| + | |
| - | <div class="col-xs-12">
| + | |
| - | <div class="project-entry-tags">
| + | |
| - | <ul>
| + | |
| - | <li><a href="#top">GO TO TOP</a></li>
| + | |
| - | </ul>
| + | |
| - | </div>
| + | |
| - | </div>
| + | |
| - | </div>
| + | |
| - |
| + | |
| - | <div class="row">
| + | |
| - | <div class="col-xs-12">
| + | |
| - | <div class="project-number">6</div><div class="project-headline-float"><h2 class="green-text project-headline"> Lab Summary</h2></div>
| + | |
| - | </div>
| + | |
| - | </div>
| + | |
| - |
| + | |
| - | <div class="row">
| + | |
| - | <!--<div class="col-sm-4 hidden-xs">
| + | |
| - | <img src="img/blog/Team_Berlin_bacteria.png" class="img-responsive" style="margin: 0 auto;"/>
| + | |
| - | </div>-->
| + | |
| - | <div class="col-xs-12 col-sm-12 blog-text" style="margin-bottom:40px;">
| + | |
| - | <p>
| + | |
| - | <a name="lab-summary"> </a>
| + | |
| - | <div class="sub-content-project">
| + | |
| - | <h2 class="sub-content-project-headline">Week 1: 02.04.2014 – 06.04.2014</h2>
| + | |
| - | Cultivation of E. coli Nissle 1917<br/>
| + | |
| - |
| + | |
| - | Genomic DNA extraction of E.coli Nissle strain<br/>
| + | |
| - |
| + | |
| - | Production of BfR, FTNA1 and FTNA2<br/>
| + | |
| - | Restriction digest<br/>
| + | |
| - | PCR Purification<br/>
| + | |
| - | Ligation Assay<br/>
| + | |
| - | Transformation into DH5α-Cells<br/>
| + | |
| - | </div>
| + | |
| - | <div class="sub-content-project">
| + | |
| - | <h2 class="sub-content-project-headline">Week 2: 07.04.2014 – 13.04.2014</h2>
| + | |
| - | Colony PCR to check the results<br/>
| + | |
| - |
| + | |
| - | Bfr, FTNA 1, FTNA 2 primar designed for amplification<br/>
| + | |
| - |
| + | |
| - | Transformation of pQE_80L into DH5α<br/>
| + | |
| - |
| + | |
| - | Cultivation of BfR, FTNA 1, FTNA 2 in LB<br/>
| + | |
| - | </div>
| + | |
| - | <div class="sub-content-project">
| + | |
| - | <h2 class="sub-content-project-headline">Week 3: 14.04.2014 – 20.04.2014</h2>
| + | |
| - | Miniprep of the cells from Week 2 <br/>
| + | |
| - |
| + | |
| - | DNA concentration determination and sequencing<br/>
| + | |
| - |
| + | |
| - | Expression of BfR, FTNA 1, FTNA 2 and induction with IPTG<br/>
| + | |
| - |
| + | |
| - | Production of a Mutaflor-Supression culture and streaking<br/>
| + | |
| - | </div>
| + | |
| - | <div class="sub-content-project">
| + | |
| - | <h2 class="sub-content-project-headline">Week 4: 21.04.2014 – 27. 04.2014 </h2>
| + | |
| - | Minirep
| + | |
| - | </div>
| + | |
| - | <div class="sub-content-project">
| + | |
| - | <h2 class="sub-content-project-headline">Week 5: 28.04.2014 – 04.05.2014</h2>
| + | |
| - | PCR Plasmid/Primar and Parameter check with Q5 Polymerase and Phusion Polymerase<br/>
| + | |
| - |
| + | |
| - | Miniprep of [pKD46+ DH5α] and [pKD4 + DH5 α]<br/>
| + | |
| - | Genomic DNA extraction from Pseudomonas putida<br/>
| + | |
| - | Enzyme digestion of different plasmids<br/>
| + | |
| - | </div>
| + | |
| - | <div class="sub-content-project">
| + | |
| - | <h2 class="sub-content-project-headline">Week 6: 05.05.2014 – 11.05.2014</h2>
| + | |
| - | Preculture of strains from Budisa strain databse in LB medium and Midiprep<br/>
| + | |
| - |
| + | |
| - | Canamycin cassette PCR of pKD4<br/>
| + | |
| - | Restriction digest of Plasmid pKD4 and pKD46v<br/>
| + | |
| - | </div>
| + | |
| - | <div class="sub-content-project">
| + | |
| - | <h2 class="sub-content-project-headline">Week 7: 12.05.2014 – 18.05.2014</h2>
| + | |
| - | Biotransformation of Ferritin and pKD46 in Nissle and DH5 α strains
| + | |
| - | </div>
| + | |
| - | <div class="sub-content-project">
| + | |
| - | <h2 class="sub-content-project-headline">Week 8: 19.05.2014 – 25.05.2014</h2>
| + | |
| - | Gel extraction of FieF PCR<br/>
| + | |
| - |
| + | |
| - | Gene Knockout of FieF in [RV + pKD46] and [WM10+pKD46]<br/>
| + | |
| - |
| + | |
| - | Transformation of pKD46 in Nissle and MG 1655<br/>
| + | |
| - |
| + | |
| - | PCR of ATPCS and PPMT<br/>
| + | |
| - |
| + | |
| - | Digestion of pQE_80L for cloning<br/>
| + | |
| - | </div>
| + | |
| - | <div class="sub-content-project">
| + | |
| - | <h2 class="sub-content-project-headline">Week 9: 26.05.2014 – 01.06.2014</h2>
| + | |
| - | Production of LB plates<br/>
| + | |
| - |
| + | |
| - | AMB-1 and Microfluidic Chip were picked up from Max-Plank Institute<br/>
| + | |
| - | </div>
| + | |
| - | <div class="sub-content-project">
| + | |
| - | <h2 class="sub-content-project-headline">Week 10: 02.06.2014 – 08.06.2014</h2>
| + | |
| - | Colony PCR and analytical gel electrophoresis for identifying the right clones<br/>
| + | |
| - |
| + | |
| - | PCR of ATPCS and PPMT to identify the correct amplification parameter.<br/>
| + | |
| - |
| + | |
| - | Colony PCR for Knockout.<br/>
| + | |
| - |
| + | |
| - | PCR of ATPCS and PPMT with Q5 High Fidelity Mastermix.<br/>
| + | |
| - | </div>
| + | |
| - | <div class="sub-content-project">
| + | |
| - | <h2 class="sub-content-project-headline">Week 11: 09.06.2014 – 15.06.2014</h2>
| + | |
| - | Refine the PCR of PPMT and ATPCS with DMSO<br/>
| + | |
| - | </div>
| + | |
| - | <div class="sub-content-project">
| + | |
| - | <h2 class="sub-content-project-headline">Week 12: 16.06.2014 – 22.06.2014</h2>
| + | |
| - | PCR amplification of ATPCS from cDNA<br/>
| + | |
| - | Colony PCR to seperate cells with FieF Knockout<br/>
| + | |
| - | Restrcition digest of pQE_80L and ATPCS-PCR Fragment<br/>
| + | |
| - | Test expression of Ferritin in RV308 (pSB1C3_Ferritin) and Nissle (pSB1C3_Ferritin)<br/>
| + | |
| - | </div>
| + | |
| - | <div class="sub-content-project">
| + | |
| - | <h2 class="sub-content-project-headline">Week 13: 23.06.2014 – 29.06.2014</h2>
| + | |
| - | Amplification of ATPCS from cDNA and PCR Purification<br/>
| + | |
| - | Preparation of Nissle, Nissle + Ferritin, RV 308, Rv 308 + Ferritin pre-cultures<br/>
| + | |
| - | Transformation of human Ferritin on PC514 to Nissle and RV 308<br/>
| + | |
| - | Induction of Ferritin expression with IPTG<br/>
| + | |
| - | </div>
| + | |
| - | <div class="sub-content-project">
| + | |
| - | <h2 class="sub-content-project-headline">Week 14: 30.06.2014 – 06.07.2014</h2>
| + | |
| - | Colony PCR and Ligation of ATPCS<br/>
| + | |
| - | Knockout of FieF and FUR in RV308 and Nissle<br/>
| + | |
| - | Transformation of RFP device and Ferritin in RV308, Nissle and WM110 strains.<br/>
| + | |
| - | PCR amplification of knockout cassette FUR/FieF<br/>
| + | |
| - | Expression of RV308 (RFP) and RV308 (RFP + Ferritin) after Induction with IPTG<br/>
| + | |
| - | </div>
| + | |
| - | <div class="sub-content-project">
| + | |
| - | <h2 class="sub-content-project-headline">Week 15: 07.07.2014 – 13.07.2014</h2>
| + | |
| - | Preparation of cryostocks and pre-cultures of ATPCS clone 1-10, WM 110, RV 308, Nissle<br/>
| + | |
| - | PCR of PPMT with goTaq Polymerase<br/>
| + | |
| - | Miniprep of ATPCS clone number 3<br/>
| + | |
| - | </div>
| + | |
| - | <div class="sub-content-project">
| + | |
| - | <h2 class="sub-content-project-headline">Week 16: 14.07.2014-20.07.2014</h2>
| + | |
| - | Midiprep of pQE_80L , PMA-T_PPMT, pKD46 <br/>
| + | |
| - | Digestion of pQE_80L with Hind III and SacI<br/>
| + | |
| - | </div>
| + | |
| - | <div class="sub-content-project">
| + | |
| - | <h2 class="sub-content-project-headline">Week 17: 21.07.2014-27.07.2014</h2>
| + | |
| - | Miniprep and Restriction of pBADex-mYFP Venus (Amp); pEx-HisII (Amp); pJS418_Phagemid (dummy) (Cm)<br/>
| + | |
| - | Ligation of PPMT Fragment from pMAC_PPMT into pEX_HisII<br/>
| + | |
| - | </div>
| + | |
| - | <div class="sub-content-project">
| + | |
| - | <h2 class="sub-content-project-headline">Week 18: 28.07.2014-3.08.2014</h2>
| + | |
| - | Degradation test of prepped TB-Expression Plasmids <br/>
| + | |
| - | Transformation of pEX_His_PPMT in MG1655, DH5α, DH10B strains<br/>
| + | |
| - | Colony PCR of the PPMT clones<br/>
| + | |
| - | Miniprep and Sequencing of pEX_His_PPMT in DH5 α<br/>
| + | |
| - | Ligation of ATPCS PCR Fragment into pQE_80 L<br/>
| + | |
| - | Colony PCR of pQE_80L_ATPCS<br/>
| + | |
| - | </div>
| + | |
| - | <div class="sub-content-project">
| + | |
| - | <h2 class="sub-content-project-headline">Week 19: 04.08.2014 -10.08.2014</h2>
| + | |
| - | Preculture of ATPCS clones for Sequencing <br/>
| + | |
| - | PCR of BamHI_PPMT_GS, GS_ATPCS_HindIII, BamHi_HuFerritin_HindIII<br/>
| + | |
| - | Transformation of BFR M52H in DH10B.<br/>
| + | |
| - | </div>
| + | |
| - | <div class="sub-content-project">
| + | |
| - | <h2 class="sub-content-project-headline">Week 20: 11.08.2014 - 17.08.2014</h2>
| + | |
| - | Prepare chemically competent cells<br/>
| + | |
| - | Generation of Heme-free BFR by Site-directed Mutagenesis<br/>
| + | |
| - | Digestion of various PCR fragements for cloning into pQE_80L<br/>
| + | |
| - | Colony PCR of pQE_80L<br/>
| + | |
| - | Clone Sequencing<br/>
| + | |
| - | </div>
| + | |
| - | <div class="sub-content-project">
| + | |
| - | <h2 class="sub-content-project-headline">Week 21: 18.08.2014 – 24.08.2014</h2>
| + | |
| - | SDS-PAGE with Coomasie staining for identification of protein expression<br/>
| + | |
| - | </div>
| + | |
| - | <div class="sub-content-project">
| + | |
| - | <h2 class="sub-content-project-headline">Week 22: 25.08.2014 – 31.08.2014</h2>
| + | |
| - | Calibration curve for iron concentration measurement<br/>
| + | |
| - | </div>
| + | |
| - | <div class="sub-content-project">
| + | |
| - | <h2 class="sub-content-project-headline">Week 23: 15.09.2014 – 21.09.2014</h2>
| + | |
| - | Preparing cultures for fluorescence microscopy<br/>
| + | |
| - | Constructing pQE_80L_T5_ATPCS_lac_PPMT<br/>
| + | |
| - | Digest and Dephosphorylation of vector pQE_80L_ATPCS<br/>
| + | |
| - | PCR of Biobrick parts (BB0-BB3)<br/>
| + | |
| - | Digest of pSB1C3-Ferritin<br/>
| + | |
| - | </div>
| + | |
| - | <div class="sub-content-project">
| + | |
| - | <h2 class="sub-content-project-headline">Week 24: 22.09.2014 – 28.09.2014</h2>
| + | |
| - | Preparation of pre-cultures<br/>
| + | |
| - | Checking insertion of gene in ATPCS_PPMT clones by colony PCR<br/>
| + | |
| - | Digestion of miniprepped pSB1C3_Ferritin (Calgary) for extraction of vector for Biobrick preparation<br/>
| + | |
| - | Mutagenesis of ATPCS in different plasmid_ATPCS construct<br/>
| + | |
| - | Berlin Vector – pQE-80L-JBFS- huFerritin Assembly PCR<br/>
| + | |
| - | Isolation of plasmid DNA of pQE80L_ATPCSMut_GS_PPMT and pQE80L_ATPCS_PPMT <br/>
| + | |
| - | Repeat PCR of Biobricks<br/>
| + | |
| - | </div>
| + | |
| - | <div class="sub-content-project">
| + | |
| - | <h2 class="sub-content-project-headline">Week 25: 29.09.2014 – 05.10.2014</h2>
| + | |
| - | Digestion of Biobrick parts<br/>
| + | |
| - | Ligation of Biobrick parts with pSB1C3 backbone<br/>
| + | |
| - | Plasmid Digestion and checked with PCR<br/>
| + | |
| - | </div>
| + | |
| - | <div class="sub-content-project">
| + | |
| - | <h2 class="sub-content-project-headline">Week 25 06.10.2014 – 12.10.2014</h2>
| + | |
| - | Colony PCR<br/>
| + | |
| - | Miniprep of cultures and preparation of sequencing samples<br/>
| + | |
| - | Preculture of knockouts<br/>
| + | |
| - | PCR of pSB1C3 backbone<br/>
| + | |
| - | Extraction of PCR Product (pSB1C3 linearised)<br/>
| + | |
| - | Iron concentration measurement in knockout strains<br/>
| + | |
| - | </div>
| + | |
| - | </div>
| + | |
| - | </div>
| + | |
| - | </div>
| + | |
| - | | + | |
| - | <!--- ////////////////// Notebook////////////////// -->
| + | |
| - | <div class="container">
| + | |
| - | <div class="row hidden-xs">
| + | |
| - | <div class="col-xs-12">
| + | |
| - | <div class="project-entry-tags">
| + | |
| - | <ul>
| + | |
| - | <li><a href="#top">GO TO TOP</a></li>
| + | |
| - | </ul>
| + | |
| - | </div>
| + | |
| - | </div>
| + | |
| - | </div>
| + | |
| - |
| + | |
| - | <div class="row">
| + | |
| - | <div class="col-xs-12">
| + | |
| - | <div class="project-number">7</div><div class="project-headline-float"><h2 class="green-text project-headline"> Notebook</h2></div>
| + | |
| - | </div>
| + | |
| - | </div>
| + | |
| - |
| + | |
| - | <div class="row">
| + | |
| - | <!--<div class="col-sm-4 hidden-xs">
| + | |
| - | <img src="img/blog/Team_Berlin_bacteria.png" class="img-responsive" style="margin: 0 auto;"/>
| + | |
| - | </div>-->
| + | |
| - | <div class="col-xs-12 col-sm-12 blog-text" style="margin-bottom:40px;">
| + | |
| - | <p>
| + | |
| - | <a name="notebook"> </a>
| + | |
| - | <div class="sub-content-project">
| + | |
| - | <h2 class="sub-content-project-headline">04/02 Wednesday</h2>
| + | |
| - | == <big><big>Cultivating E. coli Nissle 1917</big></big> ==
| + | |
| - | | + | |
| - | LB plates were produced without antibiotic. Therefore 10 ml MQ-H<sub>2</sub>O were sterile filtered in 15 ml falcon tube. 1 capsule Mutaflor 331800 was solved in the sterile water.
| + | |
| - | The E. coli Nissle solution was incubated at 37°C and 200 rpm for 1 h and crossed out at 4 plates to get different dilution of cells.
| + | |
| - | | + | |
| - | {| class="wikitable"
| + | |
| - | |-
| + | |
| - | ! dilution !! E. coli Nissle solution !! H<sub>2</sub>O !! dilution factor
| + | |
| - | |-
| + | |
| - | | I || 2 µl || 1998 µl || 1:10<sup>3</sup>
| + | |
| - | |-
| + | |
| - | | II || 100 µl from dilution I || 900 µl ||1:10<sup>4</sup>
| + | |
| - | |-
| + | |
| - | | III || 10 µl from dilution II || 990 µl || 1:10<sup>5</sup>
| + | |
| - | |}
| + | |
| - | </div>
| + | |
| - | </div>
| + | |
| - | </div>
| + | |
| - | </div>
| + | |
| - | | + | |
| - | | + | |
| - | | + | |
| - | </section> | + | |
| - | | + | |
| - | </div>
| + | |
| - | | + | |
| | | | |
| | | | |
| Line 581: |
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| | <img src="https://static.igem.org/mediawiki/2014/7/79/Team_Berlin_tuberlin_logo.png" class="img-responsive grayscale"> | | <img src="https://static.igem.org/mediawiki/2014/7/79/Team_Berlin_tuberlin_logo.png" class="img-responsive grayscale"> |
| | </div> | | </div> |
| | + | |
| | + | <div class="col-sm-2 col-sm-offset-1 col-xs-3"> |
| | + | <img src="https://static.igem.org/mediawiki/parts/c/cb/Team_Berlin_florian_renner_logo.png" class="img-responsive grayscale"> |
| | + | </div> |
| | </div> | | </div> |
| | | | |
| Line 636: |
Line 242: |
| | $(".main-menue-ul a").each(function() { | | $(".main-menue-ul a").each(function() { |
| | // checks if its the same on the address bar | | // checks if its the same on the address bar |
| - | if(url == (this.href)) { | + | if(url <strong> (this.href)) { |
| | $(this).children("li").addClass("active"); | | $(this).children("li").addClass("active"); |
| | $(this).removeClass("main-menue-links"); | | $(this).removeClass("main-menue-links"); |
"
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