ULB/13 August 2009

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== August 13, 2009 ==
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<html xmlns="http://www.w3.org/1999/xhtml">
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<div style='color:grey;'>
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==Manipulation on plasmides==
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*Les bactéries qui ont reçu le plasmide avec RBS (BBa_B0034) ont bien poussé.
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*La ligation du promoteur-RBS-Plasmide (résistant au chloramphénicol) n’a pas fonctionné : la raison pourrait être protocole l’utilisation  du manuel d’assemblage, qui nous disait de laisser liguer 10 minutes à température ambiante, puis de désactiver les enzymes alors que la notice de la T4 ligase, conseil de la laisser 24h.
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*Vérification que les bricks sont présent dans le milieu dans nos extractions : Digestion du promoteur, promoteur+RBS, RBS+RFP et le terminateur par les enzymes de restrictions EcoRI et Spel pour les 2 premiers et Xbal et Pstl pour les autres.
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*Nous avons fait une PCR avec les amorces reçues pour ccdA programme à 55°C puis migration sur gel
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==Results==
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<head>
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<meta http-equiv="content-type" content="text/html;charset=utf-8" />
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<title>iGEM Team:ULB-Brussels Wiki</title>
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<link rel="stylesheet" type="text/css" href="style.css" media="screen" />
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<link rel="stylesheet" type="text/css" href="menu.css" media="screen" />
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[[Image:13_8.jpg| les puits du dessus : Promoteur1, promoteur2, RBS+RFP1, RBS+RFP2, promoteur+RBS1, promoteur+RBS2, terminateur1, terminateur2.les puits du dessus : les résultats des migations des PCR]]
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//  -->
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        <style type="text/css">
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/* Design for https://2009.igem.org/Team:ULB-Brussels
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    Université Libre de Bruxelles
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*/
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*[[Vérification des bricks]]:4 Adn amorces « normales », blanc amorces « normales », 4 ADN amorces inverses, blanc amorces inverses.
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html body {
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width: 910px;
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font-family: Arial;
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margin: 0 auto;
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*[[PCR]]: tous les tubes ont réussi sauf que le  tube du blanc n’est pas à sa place (4eme position à la place de 6eme).
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}
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html body div.header {
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margin: 0px;
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padding: 0px;
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margin-top: -21px; /* strange... */
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}
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/* ===== banner ===== */
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html body div.header div.banner {
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height: 171px;
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background-image: url(https://static.igem.org/mediawiki/2009/0/0a/Logoulb1.png);
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html body div.header div.banner h1 {
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padding-top: 50px;
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float: left;
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width: 120px;
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margin-top: 4px;
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html body div.header div.submenu li :hover {
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color: #5F7EB4;
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color: #5F7EB4;
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/* panel = box containing mascotte, contdown and links to sponsors */
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    color: #F5B455;
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html body div.panel div.countdown {
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/* == sponsors == */
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html body div.panel div.sponsor {
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    margin-bottom: 10px;
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    min-height: 50px;
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}
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html body div.panel div.sponsor img {
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    width: 50px;
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html body div.main {
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font-size: small;
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padding-right: 10px;
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html body div.main p.abstract {
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font-weight: bold;
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html body div.main div.welcome img {
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    width: 626px;
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    height: 150px;
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    border-style: none;
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}
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html body div.footer {
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}
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</head>
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    <!-- *** Uncomment next line to enable autorefresh (developper mode) *** -->
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<!-- <body onload="JavaScript:timedRefresh(5000);"> -->
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<!-- <body> -->
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    <body>
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        <!--
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            header : box on the top of the page containing the banner and links to sections and subsections of
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            the wiki.
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        -->
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<div class="header">
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<div class="banner">
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<h1></h1>
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</div>
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<div class="menu">
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<ul>
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<li><a class="mainlink" href="/Team:ULB-Brussels">Home</a></li>
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<li><a class="mainlink" href="/Team:ULB-Brussels/Team">Team</a></li>
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<li><a class="mainlink" href="/Team:ULB-Brussels/Project">Project</a></li>
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<li><a class="mainlink" href="/Team:ULB-Brussels/Parts">Parts</a></li>
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<li><a class="mainlink" href="/Team:ULB-Brussels/Safety">Safety</a></li>
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<li><a class="mainlink selection" href="/Team:ULB-Brussels/Notebook">Notebook</a></li>
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<li><a class="mainlink" href="/Team:ULB-Brussels/Sponsors">Sponsors</a></li>
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</ul>
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<div class="submenu">
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<ul>
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<!--
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<li><a href="#english">English</a></li>
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<li><a href="#french">French</a></li>
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<li><a href="#dutch">Dutch</a></li>
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<li><a href="#german">German</a></li>
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<li><a href="#spanish">Spanish</a></li>
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<li><a href="#italian">Italiano</a></li>
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<li><a href="#chinese">Chinese</a></li>
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-->
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</ul>
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        <!--
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            panel : box on the right containing a countdown, links to the sponsors
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            [ REMOVED ]
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        <!--
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            main : main part of the page, containing the text
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<div class="footer">
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</div>
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</body>
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</html>
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<html>
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<style type="text/css">
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table.calendar          { margin: 0; padding: 10px; }
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table.calendar td      { margin: 0; padding: 2px; vertical-align: top; }
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{| align="right"
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{| style="color:#0c6;background-color:#FFFFFF;" cellpadding="3" cellspacing="1" border="1" bordercolor="#035e7b"  align="right"
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!align="right"|{{#calendar: title=ULB |year=2009 | month=07}}
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|-
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!align="right"|{{#calendar: title=ULB |year=2009 | month=08}}
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|-
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!align="right"|{{#calendar: title=ULB |year=2009 | month=09}}
 +
 +
|}
 +
 +
 +
 +
==Manipulation on plasmides==
 +
*Bacteria with the plasmid with RBS (BBa_B0034) growed.
 +
*The ligation of the promoter-RBS-plasmid (chloramphenicol resistant) didn't work: the reason could be the use of the protocol of manual assembly, who told us to leave league 10 minutes at room temperature, then deactivate the enzymes, insted of 24 hours by the T4 ligase guide.
 +
*Verify that the bricks are now in the medium of our extractions: Digestion of the promoter and promoter + RBS by restriction enzymes EcoRI and Spel, RBS + RFP and the terminator by XbaI and PstI.
 +
*We made a PCR with primers for ccdA program received at 55 ° C then a gel migration.
 +
 +
==Results==
 +
 +
[[Image:13_8.jpg| les puits du dessus  : Promoteur1, promoteur2, RBS+RFP1, RBS+RFP2, promoteur+RBS1, promoteur+RBS2, terminateur1, terminateur2.les puits du dessus : les résultats des migations des PCR]]
 +
*'''Verification of bricks''':4 DNA primers "normal", white primers "normal", 4 DNA primers reverse, white reverse primers.
 +
* '''PCR''': all tubes have succeeded except the one control sample which is out of place (4th position instead of 6th).
</div>
</div>

Latest revision as of 21:13, 21 October 2009

iGEM Team:ULB-Brussels Wiki


July
MTWTFSS
    1 2 3 4 5
6 7 8 9 10 11 12
13 14 15 16 17 18 19
20 21 22 23 24 25 26
27 28 29 30 31
August
MTWTFSS
          1 2
3 4 5 6 7 8 9
10 11 12 13 14 15 16
17 18 19 20 21 22 23
24 25 26 27 28 29 30
31
September
MTWTFSS
  1 2 3 4 5 6
7 8 9 10 11 12 13
14 15 16 17 18 19 20
21 22 23 24 25 26 27
28 29 30


Manipulation on plasmides

  • Bacteria with the plasmid with RBS (BBa_B0034) growed.
  • The ligation of the promoter-RBS-plasmid (chloramphenicol resistant) didn't work: the reason could be the use of the protocol of manual assembly, who told us to leave league 10 minutes at room temperature, then deactivate the enzymes, insted of 24 hours by the T4 ligase guide.
  • Verify that the bricks are now in the medium of our extractions: Digestion of the promoter and promoter + RBS by restriction enzymes EcoRI and Spel, RBS + RFP and the terminator by XbaI and PstI.
  • We made a PCR with primers for ccdA program received at 55 ° C then a gel migration.

Results

les puits du dessus  : Promoteur1, promoteur2, RBS+RFP1, RBS+RFP2, promoteur+RBS1, promoteur+RBS2, terminateur1, terminateur2.les puits du dessus : les résultats des migations des PCR

  • Verification of bricks:4 DNA primers "normal", white primers "normal", 4 DNA primers reverse, white reverse primers.
  • PCR: all tubes have succeeded except the one control sample which is out of place (4th position instead of 6th).

</div>

Retrieved from "http://2009.igem.org/ULB/13_August_2009"