Difference between revisions of "Team:Ionis Paris/Cloning Strategy"
Alexandra m (Talk | contribs) |
|||
| (15 intermediate revisions by 2 users not shown) | |||
| Line 7: | Line 7: | ||
<link href='https://fonts.googleapis.com/css?family=Open+Sans:400,600,700,600italic,400italic,700italic,300,300italic' rel='stylesheet' type='text/css'> | <link href='https://fonts.googleapis.com/css?family=Open+Sans:400,600,700,600italic,400italic,700italic,300,300italic' rel='stylesheet' type='text/css'> | ||
| − | + | <!-- Nos feuilles de style --> | |
| − | + | <link rel="stylesheet" type="text/css" href="https://2016.igem.org/Template:IONIS_Paris-style-css-template?action=raw&ctype=text/css" /> | |
| − | + | <!-- Version minifiée dans le bon ordre de toutes les feuilles de style du template --> | |
| + | <link rel="stylesheet" type="text/css" href="https://2016.igem.org/Template:IONIS_Paris-style-css?action=raw&ctype=text/css" /> | ||
<!-- ====BREADCUM START==== --> | <!-- ====BREADCUM START==== --> | ||
| Line 17: | Line 18: | ||
<div class="col-sm-12"> | <div class="col-sm-12"> | ||
<div class="banner_title"> | <div class="banner_title"> | ||
| − | <h1>Cloning strategy</h1> | + | <h1 id="back_to_the_top">Cloning strategy</h1> |
</div> | </div> | ||
</div> | </div> | ||
| Line 41: | Line 42: | ||
</h4> | </h4> | ||
| − | <img src="https://static.igem.org/mediawiki/2016/ | + | <img src="https://static.igem.org/mediawiki/2016/8/87/T--Ionis_Paris--Cloning_StrategyFig1.png" alt=""> |
| + | <p><i>Figure 1: Biosensor Part 1: (532bp)</i></p> | ||
<h4 class="blog_topHd"> | <h4 class="blog_topHd"> | ||
| Line 47: | Line 49: | ||
</h4> | </h4> | ||
| − | <img src="https://static.igem.org/mediawiki/2016/ | + | <img src="https://static.igem.org/mediawiki/2016/a/ac/T--Ionis_Paris--Cloning_StrategyFig2.png" alt=""> |
| + | <p><i>Figure 2: Biosensor Part 2: (1 785bp)</i></p> | ||
<h4 class="blog_topHd"> | <h4 class="blog_topHd"> | ||
| Line 53: | Line 56: | ||
</h4> | </h4> | ||
| − | <img src="https://static.igem.org/mediawiki/2016/ | + | <img src="https://static.igem.org/mediawiki/2016/7/71/T--Ionis_Paris--Cloning_StrategyFig3.png" alt=""> |
| + | <p><i>Figure 3: Biosensor Part 3: (1 215 bp)</i></p> | ||
<p></br>We began with the amplification of this parts by PCR in order to increase our stock. We used primers A12 and A13, that respectively bind the prefix and the suffix.</p> | <p></br>We began with the amplification of this parts by PCR in order to increase our stock. We used primers A12 and A13, that respectively bind the prefix and the suffix.</p> | ||
| − | <img src="https://static.igem.org/mediawiki/2016/ | + | <img src="https://static.igem.org/mediawiki/2016/1/19/T--Ionis_Paris--CloningStratTest.png" alt=""> |
| − | <p><i>Figure 4: | + | <p><i>Figure 4: Primers A12 and A13</i></p> |
</div> | </div> | ||
| Line 68: | Line 72: | ||
<img src="https://static.igem.org/mediawiki/2016/e/eb/T--Ionis_Paris--CSstep2.jpg" alt=""> | <img src="https://static.igem.org/mediawiki/2016/e/eb/T--Ionis_Paris--CSstep2.jpg" alt=""> | ||
| + | <p><i>Figure 5: Part inclusion in pSB1C3</i></p> | ||
<p>The second step was to store our 3 parts in plasmids and then bacteria. For this we digested and ligated our 3 parts in pSB1C3 (forming BB1, BB2, BB3), and transformed these biobricks in E.Coli DH5 alpha in order to amplify them. After a check of correct clones thanks to colony PCR, we purified our correct biobricks and sequenced them.</p> | <p>The second step was to store our 3 parts in plasmids and then bacteria. For this we digested and ligated our 3 parts in pSB1C3 (forming BB1, BB2, BB3), and transformed these biobricks in E.Coli DH5 alpha in order to amplify them. After a check of correct clones thanks to colony PCR, we purified our correct biobricks and sequenced them.</p> | ||
| Line 79: | Line 84: | ||
<img src="https://static.igem.org/mediawiki/2016/c/cd/T--Ionis_Paris--CSstep3.jpg" alt=""> | <img src="https://static.igem.org/mediawiki/2016/c/cd/T--Ionis_Paris--CSstep3.jpg" alt=""> | ||
| + | <p><i>Figure 6: Assembly of Biosensor Part 1 with Biosensor Part 2</i></p> | ||
<p>The third step was to assemble Part 2 with Part 1.For this we digested and ligated Part 2 and BB1, and transformed the obtained biobrick (BB12) in E.Coli DH5 alpha in order to amplified them. After a check of correct clones thanks to a PCR colony, we purified our correct biobricks and sequenced them.</p> | <p>The third step was to assemble Part 2 with Part 1.For this we digested and ligated Part 2 and BB1, and transformed the obtained biobrick (BB12) in E.Coli DH5 alpha in order to amplified them. After a check of correct clones thanks to a PCR colony, we purified our correct biobricks and sequenced them.</p> | ||
| Line 89: | Line 95: | ||
<img src="https://static.igem.org/mediawiki/2016/0/02/T--Ionis_Paris--CSstep4.jpg" alt=""> | <img src="https://static.igem.org/mediawiki/2016/0/02/T--Ionis_Paris--CSstep4.jpg" alt=""> | ||
| + | <p><i>Figure 7: Assembly of Biosensor Part 3 to the Biosensor Part 1+2</i></p> | ||
<p>The fourth step was to assemble Part 3 with Part 1 and 2. For this we digested and ligated Part 3 and BB12, and transformed the obtained biobrick (BB123) in E.Coli DH5 alpha in order to amplify them. After a check of correct clones thanks to a PCR colony, we purified our correct biobricks and sequenced them.</p> | <p>The fourth step was to assemble Part 3 with Part 1 and 2. For this we digested and ligated Part 3 and BB12, and transformed the obtained biobrick (BB123) in E.Coli DH5 alpha in order to amplify them. After a check of correct clones thanks to a PCR colony, we purified our correct biobricks and sequenced them.</p> | ||
| + | |||
| + | <p><b>Please note that all parts and primers can be found in the "Parts and Characterization" page, along with the primer sequences and the sequencing files</b></p> | ||
</div> | </div> | ||
| Line 205: | Line 214: | ||
<!-- ====END BLOG TABLE==== --> | <!-- ====END BLOG TABLE==== --> | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | <!-- ====START SOCIAL Link==== --> | |
| − | + | <div class="footer_social"> | |
| − | + | <div class="container-fluid"> | |
| − | + | <div class="row"> | |
| − | + | <ul class="ft_top clearfix"> | |
| − | + | <li class="waves-effect waves-light"> | |
| − | + | <a href="https://www.facebook.com/ionisigem/?fref=ts" target="_blank"> | |
| − | + | <i class="zmdi zmdi-facebook "></i>Facebook | |
| − | + | </a> | |
| − | + | </li> | |
| − | + | <li class="waves-effect waves-light"> | |
| − | + | <a href="https://twitter.com/igem_ionis" target="_blank"> | |
| − | + | <i class="zmdi zmdi-twitter"></i>Twitter | |
| − | + | </a> | |
| − | + | </li> | |
| − | + | <li class="waves-effect waves-light"> | |
| − | + | <a href="https://www.youtube.com/channel/UC0RyQsB5YpweSRBYQlAdZDA" target="_blank"> | |
| − | + | <i class="zmdi zmdi-youtube"></i>youtube | |
| − | + | </a> | |
| − | + | </li> | |
| − | + | <li class="waves-effect waves-light"> | |
| − | + | <a href="https://www.linkedin.com/company/igem-ionis" target="_blank"> | |
| − | + | <i class="zmdi zmdi-linkedin"></i>LinkedIn | |
| − | + | </a> | |
| − | + | </li> | |
| − | + | ||
| − | + | </ul> | |
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | </ | + | |
</div> | </div> | ||
</div> | </div> | ||
| − | + | </div> | |
| − | + | <!-- ====END FOOTER TOP==== --> | |
| − | + | ||
| − | + | <!-- ====START FOOTER AREA==== --> | |
| − | + | <footer class="footer_area"> | |
| − | + | <div class="footer_middle"> | |
| − | + | <div class="container"> | |
| − | + | <div class="row"> | |
| − | + | <div class="col-xs-12"> | |
| − | + | <div class="scroll_area"> | |
| + | <div class="sroll_top"> | ||
| + | <a href="#ancre"> <i class="zmdi zmdi-chevron-up btn waves-effect"> </i> </a> | ||
</div> | </div> | ||
</div> | </div> | ||
</div> | </div> | ||
| + | </div> | ||
| + | <div class="row"> | ||
<div class="row"> | <div class="row"> | ||
<div class="col-lg-5 col-md-5 col-sm-12"> | <div class="col-lg-5 col-md-5 col-sm-12"> | ||
<div class="middle_content"> | <div class="middle_content"> | ||
| − | <h4> | + | <h4>iGEM IONIS</h4> |
| − | <p> | + | <p> We're a group of six different schools from the IONIS Education Group. For this |
| − | + | competition we wanted to take advantage of the multiple schools and fields of activity | |
| − | + | given by the IONIS education group to create a solid project.</p> | |
| + | <a href="https://2016.igem.org/Team:Ionis_Paris/Team">Read More</a> | ||
</div> | </div> | ||
</div> | </div> | ||
| + | |||
<div class="col-lg-3 col-lg-offset-1 col-md-4 col-sm-7"> | <div class="col-lg-3 col-lg-offset-1 col-md-4 col-sm-7"> | ||
<div class="footer_Widgets"> | <div class="footer_Widgets"> | ||
| Line 272: | Line 280: | ||
<ul> | <ul> | ||
<li><i class="zmdi zmdi-pin"></i> | <li><i class="zmdi zmdi-pin"></i> | ||
| − | <span>Location: | + | <span>Location: 66 Rue Guy Môquet, 94800 Villejuif, France</span> |
| − | + | ||
| − | + | ||
| − | + | ||
</li> | </li> | ||
| + | |||
<li><i class="zmdi zmdi-email"></i> | <li><i class="zmdi zmdi-email"></i> | ||
| − | <a href="mailto: | + | <a href="mailto:igem.ionis@gmail.com">email: igem.ionis@gmail.com</a> |
| − | + | ||
| − | + | ||
| − | + | ||
</li> | </li> | ||
| + | |||
</ul> | </ul> | ||
</div> | </div> | ||
</div> | </div> | ||
</div> | </div> | ||
| − | + | <div class="col-lg-3 col-md-3 col-sm-5"> | |
<div class="footer_Widgets"> | <div class="footer_Widgets"> | ||
| − | <h4> | + | <h4>Download the app</h4> |
| − | <figure class="widget_gallery"> | + | <a href="https://itunes.apple.com/us/app/quantifly/id1166875690?ls=1&mt=8" |
| − | + | target="_blank" style="target-new: tab;"> | |
| − | + | <figure class="widget_gallery"> | |
| − | + | <div class="RXcircleEffect"></div> | |
| − | < | + | <img src="https://static.igem.org/mediawiki/2016/8/8d/IONIS_IGEM_paris_logo_apple.png" |
| + | alt="" /> | ||
| + | <figcaption> | ||
<i class="zmdi zmdi-link"></i> | <i class="zmdi zmdi-link"></i> | ||
| − | </ | + | </figcaption> |
| − | + | </figure> | |
| − | + | </a> | |
| − | + | <a href="https://play.google.com/store/apps/details?id=fr.plnech.quantifly&hl=en" | |
| − | + | target="_blank" style="target-new: tab;"> | |
| − | + | <figure class="widget_gallery"> | |
| − | + | <div class="RXcircleEffect"></div> | |
| − | + | <img src="https://static.igem.org/mediawiki/2016/6/65/IONIS_IGEM_paris_google_play.png" | |
| − | + | alt="" /> | |
| − | + | <figcaption> | |
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
<i class="zmdi zmdi-link"></i> | <i class="zmdi zmdi-link"></i> | ||
| − | </ | + | </figcaption> |
| − | </ | + | </figure> |
| − | </ | + | </a> |
| + | |||
| + | |||
</div> | </div> | ||
</div> | </div> | ||
| + | |||
</div> | </div> | ||
| + | |||
</div> | </div> | ||
</div> | </div> | ||
| Line 355: | Line 331: | ||
<div class="col-md-7 fix_p_l"> | <div class="col-md-7 fix_p_l"> | ||
<nav> | <nav> | ||
| − | <ul class="ft_bottom"> | + | <ul class="ft_bottom"> |
| − | <li><a href=" | + | <li><a href="https://2016.igem.org/Team:Ionis_Paris">Home</a></li> |
| − | <li><a href=" | + | <li><a href="https://2016.igem.org/Team:Ionis_Paris/Working_at_La_Paillasse">in the Lab</a></li> |
| − | <li><a href=" | + | <li><a href="https://2016.igem.org/Team:Ionis_Paris/Measurement"> Side Projects</a></li> |
| − | <li><a href=" | + | <li><a href="https://2016.igem.org/Team:Ionis_Paris/Parts">Results</a></li> |
| − | <li><a href=" | + | <li><a href="https://2016.igem.org/Team:Ionis_Paris/HPintro">Human |
| − | <li><a href=" | + | Practice</a></li> |
| + | <li><a href="https://2016.igem.org/Team:Ionis_Paris/Team">Team</a></li> | ||
</ul> | </ul> | ||
</nav> | </nav> | ||
| Line 367: | Line 344: | ||
<div class="col-md-5 fix_p"> | <div class="col-md-5 fix_p"> | ||
<div class="ft_paragraph"> | <div class="ft_paragraph"> | ||
| − | <p> | + | <p>©IONIS_IGEM_2016</a>.</p> |
</div> | </div> | ||
| − | </div> | + | </div> |
</div> | </div> | ||
</div> | </div> | ||
</div> | </div> | ||
| − | </div> | + | </div> |
| − | + | </footer> | |
| − | + | <!-- ====END FOOTER AREA==== --> | |
| − | + | ||
| − | + | <!-- ====Google Maps API==== --> | |
| − | + | <script src="https://maps.googleapis.com/maps/api/js"></script> | |
| − | + | ||
| − | + | <!-- Nos scripts --> | |
| + | <script type="text/javascript" src="https://2016.igem.org/wiki/index.php?title=Template:IONIS_PARIS_JS&action=raw&ctype=text/javascript"></script> | ||
| + | |||
</html> | </html> | ||
Latest revision as of 20:54, 19 October 2016
In order to obtain synthesized DNA fragments without error, we chose to synthesis our big biosensor part of 3,305 bp in 3 smaller parts between 500 bp and 1,800 bp that we planned to then assemble together in the pSB1C3 backbone. Figure 1: Biosensor Part 1: (532bp) Figure 2: Biosensor Part 2: (1 785bp) Figure 3: Biosensor Part 3: (1 215 bp) We began with the amplification of this parts by PCR in order to increase our stock. We used primers A12 and A13, that respectively bind the prefix and the suffix. Figure 4: Primers A12 and A13 Figure 5: Part inclusion in pSB1C3 The second step was to store our 3 parts in plasmids and then bacteria. For this we digested and ligated our 3 parts in pSB1C3 (forming BB1, BB2, BB3), and transformed these biobricks in E.Coli DH5 alpha in order to amplify them. After a check of correct clones thanks to colony PCR, we purified our correct biobricks and sequenced them. Figure 6: Assembly of Biosensor Part 1 with Biosensor Part 2 The third step was to assemble Part 2 with Part 1.For this we digested and ligated Part 2 and BB1, and transformed the obtained biobrick (BB12) in E.Coli DH5 alpha in order to amplified them. After a check of correct clones thanks to a PCR colony, we purified our correct biobricks and sequenced them. Figure 7: Assembly of Biosensor Part 3 to the Biosensor Part 1+2 The fourth step was to assemble Part 3 with Part 1 and 2. For this we digested and ligated Part 3 and BB12, and transformed the obtained biobrick (BB123) in E.Coli DH5 alpha in order to amplify them. After a check of correct clones thanks to a PCR colony, we purified our correct biobricks and sequenced them. Please note that all parts and primers can be found in the "Parts and Characterization" page, along with the primer sequences and the sequencing files
Step 1: Synthesis of DNA parts
Part 1 (532 bp)
Part 2 (1,785 bp)
Part 3 (1,215 bp)
Step 2: Storage of our 3 parts in pSB1C3 => BB1, BB2, BB3
Step 3: Assembling of BB1 with P2 => BB12
Step 4: Assembling of BB12 with P3 => BB123 (Whole Biosensor)
