Difference between revisions of "Team:Ionis Paris/21 07 16"
Alexandra m (Talk | contribs) |
|||
| Line 166: | Line 166: | ||
<!-- ====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 advantages of the multiple schools and activity field | |
| − | + | 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 233: | Line 229: | ||
<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> | </div> | ||
| + | |||
</div> | </div> | ||
</div> | </div> | ||
| Line 316: | Line 252: | ||
<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 328: | Line 265: | ||
<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> | ||
Revision as of 12:06, 18 October 2016
The objective is to transforme competent DH5⍺ cells with the ligation products BB1, BB2 and BB3 to create a stock of transformed bacteria. 6 aliquots of NEB DH5⍺ Competent E.coli (C2287) Plasmid DNA : Ligation product BB1, BB2 and BB3 (from the 20/07/16) Petri dish LB+Cm: Cm concentration = 25 µg/mL We need 20 LB+Cm plates + 11 LB plates haw 2 tubes of DH5⍺ competent cells on ice for 10 min. Mix gently and carefully pipette 50 µL of cells into the 4 transformation tubes on ice. Add the 20 µL / 30 µL plasmid DNA to the cell mixture. Carefully flick the tubes 4-5 times to mix cells and DNA. Do not vortex. Place on ice for 30 min. Do not mix. Heat shock at exactly 42°C for 45 s. Do not mix. Place on ice for 5 min. Do not mix. Pipette 250 µL of room temperature SOC into the mixture. Place at 37°C for 1h at 250 rpm. Warm selection plates to 25°C. Mix the cells thoroughly by flicking the tubes and inverting. Spread the corresponding volume onto each plate. Incubate all the plates O/N at 37°C. Some colonies on the petri dishes LB+Cm plated with 50 µL of bacteria transformed with the different ligation products and more on the petri dishes LB+Cm plated with 200 µL of bacteria. We obtained expected results for petri-dishes plated with bacteria transformed with BB1 and BB2, but there is no colonies for petri-dishes plated with bacteria transformed with BB3. The transformation worked for BB1 and BB2. Colonies contain a plasmid with the Chloramphenicol resistance gene, present in pSB1C3. A PCR colonie is necessary to check the size of the plasmid present in colonies, and therefore in order to know if bacteria incorpore the correct plasmids BB1 and BB2. A new digestion, ligation and transformation is necessary for BB3.
Transformation: competent DH5⍺ cells with ligation products BB1, BB2 and BB3
Objectives
Materials
Protocol
Experimental conditions achieved :
Transformations protocol:
Results (obtain the 22/07)
Expected results:
A bacterial lawn on the LB petri dish without antibiotic.
No colonies on the LB+Cm petri dish plated with bacteria transformed with no plasmid (- control).Obtained results:
Interpretation
