Difference between revisions of "Team:Ionis Paris/17 07 16"
AlexDOLLET (Talk | contribs) (Created page with "{{IONIS_HEADER}} <html> <!-- ====Google font==== --> <link href='https://fonts.googleapis.com/css?family=Merriweather:400,700,700italic,400italic,300italic...") |
Alexandra m (Talk | contribs) |
||
| (5 intermediate revisions by 4 users not shown) | |||
| Line 1: | Line 1: | ||
{{IONIS_HEADER}} | {{IONIS_HEADER}} | ||
<html> | <html> | ||
| − | |||
| − | |||
| − | + | <!-- 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" /> | |
| − | + | <link rel="stylesheet" type="text/css" href="https://2016.igem.org/Template:IONIS_Paris-style-css?action=raw&ctype=text/css" /> | |
| − | + | ||
| − | + | ||
| − | + | ||
| Line 34: | Line 29: | ||
<div class="col-xs-12 col-sm-9"> | <div class="col-xs-12 col-sm-9"> | ||
<div class="bloggrid_right"> | <div class="bloggrid_right"> | ||
| − | + | ||
| − | < | + | <h2 class="blog_topHd"> <font color =”#279AD3”>Mini-prep: on DH5⍺ transformed with pSB1C3-RFP</font></h2 |
| − | + | ||
| − | + | <h3><font color =”94FAF1”> Objectives </font></h3> | |
| − | + | ||
<p>Purification and quantification of the pSB1C3-RFP plasmid extracted from the bacterial mini-cultures in order to make stock of pSB1C3-RFP plasmid.</p> | <p>Purification and quantification of the pSB1C3-RFP plasmid extracted from the bacterial mini-cultures in order to make stock of pSB1C3-RFP plasmid.</p> | ||
| − | + | <h3><font color =”94FAF1”> Materials </font></h3> | |
<p>4 mini-cultures of bacteria transformed with pSB1C3-RFP realized the 16/07 (put a colony with satisfying PCR results in 5 mL LB+CHL into a 50 mL Falcon tube).<br/> | <p>4 mini-cultures of bacteria transformed with pSB1C3-RFP realized the 16/07 (put a colony with satisfying PCR results in 5 mL LB+CHL into a 50 mL Falcon tube).<br/> | ||
| Line 47: | Line 41: | ||
| − | + | <h3><font color =”94FAF1”> Protocol </font></h3> | |
| − | <p>The miniprep were realized using the QIAprep® Spin Miniprep Kit (Qiagen, ref: 27104) and following the protocol given by the supplier</p> | + | <p>The miniprep were realized using the QIAprep® Spin Miniprep Kit (Qiagen, ref: 27104) and following the protocol given by the supplier (available <a href="https://www.qiagen.com/fr/resources/resourcedetail?id=331740ca-077f-4ddd-9e5a-2083f98eebd5&lang=en">here</a>)</p> |
| − | + | <h5><font color =”#3CB5E1”>Miniprep:</font></h5> | |
<ol> <li><p>Divide each 4.5 mL bacterial O/N mini-cultures into 4 Eppendorf tubes and centrifuge all those tubes at 9,000 rpm for 3 min at room temperature. Discard the supernatant.</p></li> | <ol> <li><p>Divide each 4.5 mL bacterial O/N mini-cultures into 4 Eppendorf tubes and centrifuge all those tubes at 9,000 rpm for 3 min at room temperature. Discard the supernatant.</p></li> | ||
| Line 66: | Line 60: | ||
<li><p>Store the purified DNA at -20°C.</p></li> | <li><p>Store the purified DNA at -20°C.</p></li> | ||
</ol> | </ol> | ||
| − | + | <h5><font color =”#3CB5E1”>Bacteria storage:</font></h5> | |
<ol> <li><p> Add 100 µL of glycerol to 100 µL of transformed bacteria in clean microcentrifuge 1.5 mL Eppendorf.</p> | <ol> <li><p> Add 100 µL of glycerol to 100 µL of transformed bacteria in clean microcentrifuge 1.5 mL Eppendorf.</p> | ||
| Line 75: | Line 69: | ||
</li> | </li> | ||
</li> | </li> | ||
| − | </ol> | + | </ol> |
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | <h3><font color =”94FAF1”> Results </font></h3> | |
| − | + | ||
| − | + | ||
| + | <h5><font color =”#3CB5E1”>Nanodrop quantification:</font></h5> | ||
| − | <figure | + | <figure> |
| − | <img src="https://static.igem.org/mediawiki/2016/7/75/T--Ionis_Paris--Notebook17_07Fig.1.png" alt=""> | + | <center><img src="https://static.igem.org/mediawiki/2016/7/75/T--Ionis_Paris--Notebook17_07Fig.1.png" alt=""></center> |
</figure> | </figure> | ||
| Line 177: | Line 150: | ||
<!-- ====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 244: | Line 213: | ||
<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> |
| − | <figure class="widget_gallery"> | + | <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 327: | Line 264: | ||
<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 339: | Line 277: | ||
<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 20:01, 19 October 2016
Purification and quantification of the pSB1C3-RFP plasmid extracted from the bacterial mini-cultures in order to make stock of pSB1C3-RFP plasmid. 4 mini-cultures of bacteria transformed with pSB1C3-RFP realized the 16/07 (put a colony with satisfying PCR results in 5 mL LB+CHL into a 50 mL Falcon tube). The miniprep were realized using the QIAprep® Spin Miniprep Kit (Qiagen, ref: 27104) and following the protocol given by the supplier (available here) Divide each 4.5 mL bacterial O/N mini-cultures into 4 Eppendorf tubes and centrifuge all those tubes at 9,000 rpm for 3 min at room temperature. Discard the supernatant. Resuspend the pellet in 62.5 μL Buffer P1 and pool the 4 Eppendorf tubes into a unique tube. Add 250 μL Buffer P2 and mix by inverting the tube 6 times. The solution turns blue. Add 350 μL Buffer N3 and mix by inverting the tube 6 times. The solution turns colorless. Centrifuge for 10 min at 13,000 rpm. Load 800 μL supernatant from step 5 to the QIAprep 2.0 spin column. Centrifuge for 1 min and discard the flow-through. Add 500 µL Buffer PB. Centrifuge for 1 min at 13,000 rpm and discard the flow-through. Add 750 µL Buffer PE. Centrifuge for 1 min at 13,000 rpm and discard the flow-through. Centrifuge once more for 1 min at 13,000 rpm. Place the QIAprep 2.0 spin column in a clean 1.5 mL microcentrifuge tube. Add 50 μL Buffer EB to the center of the QIAprep 2.0 spin column, let stand for 1 min, and centrifuge for 1 min at 13,000 rpm. Calculate the quantity of DNA with the Nanodrop. Store the purified DNA at -20°C. Add 100 µL of glycerol to 100 µL of transformed bacteria in clean microcentrifuge 1.5 mL Eppendorf. 5 tubes of pSB1C3-RFP (1-4) Store at -80°C.
Mini-prep: on DH5⍺ transformed with pSB1C3-RFP
Objectives
Materials
From those mini-cultures, take 500 µL to realize a glycerol stock of tranformed bacteria. The 4.5 mL remaining will serve for the miniprep. Protocol
Miniprep:
Bacteria storage:
Results
Nanodrop quantification:
