Difference between revisions of "Team:Bordeaux/Template:AchievementsProtocols"

Latest revision as of 17:11, 28 August 2015

Protocols

Our Organisms of Choice is Escherichia coli and Saccharomyces cerevisiae . Listed below are the protocols we used in our lab.

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-<section align="center">
+<section align="left">
      <div class="content-wrapper">
-         <div class="col-lg-10 col-lg-offset-1">
+         <div class="col-lg-10 col-lg-offset-1"align="left">
-            <h2> Protocols </h2>
-<h5> <b> Making DMSO Competent Cells </b> </h5>
+ <h3 align="center"> Protocols </h3>
-<table cellspacing="0" cellpadding="0" border="1" bgcolor="red" bordercolor="black" width="80%" align="center">
+<p> Our Organisms of Choice is <I>Escherichia coli</I> and <I>Saccharomyces cerevisiae </I>. Listed below are the protocols we used in our lab. </p>
-     <caption-top> <b><u>SOB : 1L</b></u></caption>
-     <thead> <!--En-tête du tableau -->
+<h5> Cloning </h5>
-     <tr>
-          <th>Final concentration</th>
-          <th>Composent</th>
-          <th>Volume & Mass</th>
-     </tr>
-     </thead>
-     <tbody> <!--Cors du tableau-->
+   <ul>
-     <tr>
+      <li><a href="https://static.igem.org/mediawiki/2015/b/b9/PCR_iGEM_Bordeaux_Protocol.pdf" target="_blank"><font size="3" ><B>PCR Methods iGEM Bordeaux</B></font></a></li>
-          <td>2%</td>
+      <li><a href="https://static.igem.org/mediawiki/2015/a/a3/E.coli_Competent_Cells_v2.pdf" target="_blank"><font size="3" ><B><I>E.coli</I> Competent Cells</B></font></a></li>
-          <td>Bactotryptone</td>
-          <td>20g</td>
+<li><a href="https://static.igem.org/mediawiki/2015/b/ba/Miniprep_Camilla_V2_.pdf"target="_blank"><font size="3" ><B>Plasmid Extraction using Alkaline Lysis Method</B></font></a></li>
-     </tr>
-     <tr>
+   </ul>
-          <td>0.5%</td>
-          <td>Yeast Extract</td>
-          <td>5g</td>
-     </tr>
-      <tr>
+<h5> Working with <I>E.coli</I> </h5>
-          <td>10mM</td>
-          <td>NaCl</td>
-          <td>2mL (5M stock)</td>
-     </tr>
-     <tr>
+<ul>
-          <td>2.5mM</td>
+    <li><a href="https://static.igem.org/mediawiki/2015/6/60/Growing_Medium_of_E.coli_VC.pdf"target="_blank"><font size="3" ><B>Growing Media of <I>E.coli</I></B></font></a>  </li>
-          <td>KCl</td>
-          <td>2.5mL (1M stock)</td>
-     </tr>
-      <tr>
+ <li><a href="https://static.igem.org/mediawiki/2015/0/08/E.coli_Curdlan_Purification_V%2A%2A.pdf"target="_blank"><font size="3" ><B><I>E.coli</I> Curdlan Purification</B></font></a>  </li>
-          <td>10mM</td>
-          <td>MgCl2</td>
-          <td>10mL (1M stock)</td>
-     </tr>
-      <tr>
-          <td>10mM</td>
-          <td>MgSO4</td>
-          <td>10mL (1M stock)</td>
-     </tr>
-     <tr>
-     <td colspan="3">pH media o 7 with NaOH, then autoclave</td>
-     </tr>
-     </tbody>
-</table>
-<br>
-<p align="left">- Transformation Broth (TB) : 1L</p>
+</ul>
-<!--Tableau TB à insérer -->
-<p align="left">1) Mix the Pipes, CaCl2, and KCl in 900 ml of millipore water.
+<h5> Working with <I>S.cerevisiae </I> </h5>
-<br> 2) Add NaOH until pH is 6.7 (Don’t worry, dust disappear after pH adjust)
-<br> 3) Add MnCl2 (see above), stir, adjust volume to 1 L
-<br> 4) Filter sterilize (Filters are in a cardboard box below the BET bench)
-<br> 5) Store at 4C. 1L</p>
-<br>
-<p align="left"> <b>DAY ONE :</b>
+   <ul>
-<br> 1) Grow 12 ml overnight culture of favorite strain of E. coli in 2XTY (preheat medium at 37°C before inoculation)
-<br> 2) Make SOB and TB. </p>
+    <li><a href="https://static.igem.org/mediawiki/2015/5/5a/High_Efficiency_Yeast_Transformation_.pdf"target="_blank"><font size="3" ><B>High Efficiency Yeast Transformation</B></font></a>  </li>
-<p align="left"> <b>DAY TWO :</b>
+   <li><a href="https://static.igem.org/mediawiki/2015/7/7d/Genomic_DNA_preparation_from_Yeast_.pdf"target="_blank"><font size="3" ><B> Genomic DNA Preparation from Yeast</B></font></a>  </li>
-<br> 1) Keep 5mL of SOB for initial OD.
-<br> 2) Inoculate 1 L SOB with 12 ml overnight culture.
-<br> 3) Grow culture at 18C (this temperature is really important as we see a 10-fold decrease in competency when we
-grow them at room temperature). </p>
+ </ul>
+<h5> Curdlan Methods </h5>
+ <ul>
+    <li><a href="https://static.igem.org/mediawiki/2015/b/bb/Curdlan_Quantification.pdf"target="_blank"><font size="3" ><B> Curdlan Quantification</B></font></a></li>
+    <li><a href="https://static.igem.org/mediawiki/2015/4/43/Sulfation_of_polysaccharides_by_Sodium_Pyrosulfate_in_Dim%C3%A9thylsulfoxide_VC.pdf"target="_blank"><font size="3" ><B> Sulfation of Curdlan by Sodium Pyrosulfate in Dimethyl Sulfoxide</B></font></a></li>
+</ul>
-<p align="left"> <b>DAY THREE :</b>
-<br> 1) Grow cells until A600 0.5-0.7.
-<br>
-<br> <i>Subsequent steps should be carried out in the cold room on ice:</i>
-<br> 2) Put flask on ice for 10 minutes, then spin cells down at 2500xg (3350 RPM) in JLA 8.1 rotor
-<br> 3) Pour off supernatant
-<br> 4) Resuspend gently first in 25 ml TB, then add remaining 295 ml (Final 320mL)
-<br> 5) Leave on ice for 5 minutes
-<br> 6) Spin down again at 2500xg for 10 minutes
-<br> 7) Resuspend cells in 40 ml TB
-<br> 8) Add 3 ml of DMSO dropwise while gently shaking [final DMSO concentration is 7%].
-<br> 9) Aliquot in 100 μl aliquots (you will need about 450 pre-chilled 0.5 ml tubes).
-<br> 10) Flash freeze in liquid nitrogen.
-<br> 11) Store at -80C (the lower the rack the better). </p>
-<!--Savy : Rédaction de la page 2 du protocole en cours ; mise en place des tableaux à faire (je dois plancher sur le code pour les faire aussi jolie que sur le pdf de Jean!-->
-<h5> <b> Selective Medium for Transformation </b> </h5>
-<!-- Tableau de composition à insérer seulement !-->
         </div>

Difference between revisions of "Team:Bordeaux/Template:AchievementsProtocols"

Latest revision as of 17:11, 28 August 2015

Protocols

Cloning

Working with E.coli

Working with S.cerevisiae

Curdlan Methods