Difference between revisions of "Team:Pasteur Paris/Week 14"

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               <span class="soustitrepage">08/31 - 09/04</span></p>
 
               <span class="soustitrepage">08/31 - 09/04</span></p>
 
                 <p></p><br><br><br>
 
                 <p></p><br><br><br>
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              <p style="text-align: justify; padding: 30px;">Lorem ipsum dolor sit amet, consectetur adipiscing elit. Donec et purus vel risus iaculis tristique. Ut sit amet eros mi. Nulla elementum aliquam nibh. Praesent commodo, leo sit amet luctus gravida, dolor orci fermentum dolor, et consequat velit lacus quis ligula. Cum sociis natoque penatibus et magnis dis parturient montes, nascetur ridiculus mus. Fusce ut euismod odio. Vestibulum sed congue velit. Donec vel mi eget felis elementum interdum quis ac nisi.<br/><br/>
+
             
 +
<p style="text-align:left">1/ After 72h of incubation, only contamination was  observed.</p>
 +
<p style="text-align:left"> 2/ Co-transfection in yeast of nicked pRS415 nicked and Cre with extensions the  protocol given by Heloise MULLER. </p>
 +
<p style="text-align:left">3/ After Four days of culture of the  electroporated yeast BY4742 with Cre+ homologous extension and the plasmid  pRS415 nicked as vector no colonies has grown on Petri dishes. </p>
 +
<br>
 +
<p style="text-align:left">Assembly of Cluster 1 :</p>
 +
<ul style="text-align:left">
 +
  <li>Xba I and Pst I enzymatic  digest. </li>
 +
  <li>Gel migration on 1% agarose  gel. </li>
 +
  <li>Gel Extraction following the  Macherey-Nagel Protocol </li>
 +
  <li>DNA Concentration </li>
 +
</ul>
 +
<table border="1" cellspacing="0" cellpadding="0" width="604">
 +
  <tr>
 +
    <td width="101" valign="top"><p>Dilution 1/25</p></td>
 +
    <td width="101" valign="top"><p>DNA Concentration (ng/µl)</p></td>
 +
    <td width="101" valign="top"><p>OD(230nm)</p></td>
 +
    <td width="101" valign="top"><p>OD(260nm)</p></td>
 +
    <td width="101" valign="top"><p>OD(280nm)</p></td>
 +
    <td width="101" valign="top"><p>OD(260nm)/OD(230nm)</p></td>
 +
  </tr>
 +
  <tr>
 +
    <td width="101" valign="top"><p>BBa_808030</p></td>
 +
    <td width="101" valign="top"><p align="right">0,2 </p></td>
 +
    <td width="101" valign="top"><p align="right">0,674 </p></td>
 +
    <td width="101" valign="top"><p align="right">0,04 </p></td>
 +
    <td width="101" valign="top"><p align="right">0,00 </p></td>
 +
    <td width="101" valign="top"><p align="right">0,01 </p></td>
 +
  </tr>
 +
  <tr>
 +
    <td width="101" valign="top"><p>BBa_808031</p></td>
 +
    <td width="101" valign="top"><p align="right">0,2 </p></td>
 +
    <td width="101" valign="top"><p align="right">0,560 </p></td>
 +
    <td width="101" valign="top"><p align="right">0,003 </p></td>
 +
    <td width="101" valign="top"><p align="right">0 </p></td>
 +
    <td width="101" valign="top"><p align="right">0,01 </p></td>
 +
  </tr>
 +
</table>
 +
<ul style="text-align:left">
 +
  <li>Midiprep of BBa_K808030</li>
 +
  <li>Spe I and PstI enzymatic  digest of BBa_K808007 with the phosphatase step. </li>
 +
  <li>Gel migration on 1% agarose  gel. </li>
 +
  <li>Gel Purification following  the Macherey-Nagel Protocol </li>
 +
  <li>DNA concentration assay  using a spectrophotometer. </li>
 +
</ul>
 +
<table border="1" cellspacing="0" cellpadding="0" width="604">
 +
  <tr>
 +
    <td width="86" valign="top"><p>Dilution 1/25</p></td>
 +
    <td width="86" valign="top"><p>DNA Concentration (ng/µl)</p></td>
 +
    <td width="86" valign="top"><p>OD(230nm)</p></td>
 +
    <td width="86" valign="top"><p>OD(260nm)</p></td>
 +
    <td width="86" valign="top"><p>OD(280nm)</p></td>
 +
    <td width="86" valign="top"><p>OD(260nm)/OD(230nm)</p></td>
 +
    <td width="86" valign="top"><p>OD(260nm)/OD(280nm</p></td>
 +
  </tr>
 +
  <tr>
 +
    <td width="86" valign="top"><p>BBa_808007</p></td>
 +
    <td width="86" valign="top"><p align="right">0,8 </p></td>
 +
    <td width="86" valign="top"><p align="right">0,07 </p></td>
 +
    <td width="86" valign="top"><p align="right">0,017 </p></td>
 +
    <td width="86" valign="top"><p align="right">0,014 </p></td>
 +
    <td width="86" valign="top"><p align="right">0,24 </p></td>
 +
    <td width="86" valign="top"><p align="right">1,25 </p></td>
 +
  </tr>
 +
  <tr>
 +
    <td width="86" valign="top"><p>BBa_808007</p></td>
 +
    <td width="86" valign="top"><p align="right">1,1 </p></td>
 +
    <td width="86" valign="top"><p align="right">0,133 </p></td>
 +
    <td width="86" valign="top"><p align="right">0,022 </p></td>
 +
    <td width="86" valign="top"><p align="right">0,017 </p></td>
 +
    <td width="86" valign="top"><p align="right">0,17 </p></td>
 +
    <td width="86" valign="top"><p align="right">1,28 </p></td>
 +
  </tr>
 +
</table>
 +
<ul style="text-align:left">
 +
  <li>Ligation of BBa_K808007 and  BBa_K808030 </li>
 +
  <li>Transformation in  electro-competent DH5-⍺ cells. </li>
 +
  <li>Preparation for sequencing. </li>
 +
</ul>
 +
<br>
 +
<p style="text-align:left">Assembly of cluster 2 :</p>
 +
<ul>
 +
  <li>PCR amplification of BBa_K936011  and BBa_K936023 using Q5 High fidelity Master Mix. </li>
 +
</ul>
  
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Quisque scelerisque sollicitudin convallis. In non semper dui. Proin eu dignissim justo. Suspendisse dictum imperdiet ante quis rhoncus. Nunc mollis tempor orci eget elementum. Morbi sit amet massa eleifend, egestas lectus nec, condimentum ligula. Pellentesque sit amet scelerisque ipsum.<br/><br/>
 
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Quisque posuere ex sed tortor pulvinar finibus dignissim ac lorem. Vivamus quis ullamcorper odio. Mauris ut semper mi. Phasellus metus nisi, tempor id dolor ac, condimentum scelerisque odio. Vivamus ultricies pellentesque metus. Etiam nec metus felis. Nullam facilisis sit amet ante gravida laoreet. Etiam vulputate commodo semper. Curabitur a rutrum est, id bibendum sem. Sed congue semper mauris, sed laoreet erat pulvinar non.<br/><br/>
 
 
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Revision as of 16:43, 18 September 2015

Week 1

Week 2

Week 3

Week 4

Week 5

Week 6

Week 7

Week 8

Week 9

Week 10

Week 11

Week 12

Week 13

Week 14

Week 15

Week 14

08/31 - 09/04




1/ After 72h of incubation, only contamination was observed.

2/ Co-transfection in yeast of nicked pRS415 nicked and Cre with extensions the protocol given by Heloise MULLER.

3/ After Four days of culture of the electroporated yeast BY4742 with Cre+ homologous extension and the plasmid pRS415 nicked as vector no colonies has grown on Petri dishes.


Assembly of Cluster 1 :

  • Xba I and Pst I enzymatic digest.
  • Gel migration on 1% agarose gel.
  • Gel Extraction following the Macherey-Nagel Protocol
  • DNA Concentration

Dilution 1/25

DNA Concentration (ng/µl)

OD(230nm)

OD(260nm)

OD(280nm)

OD(260nm)/OD(230nm)

BBa_808030

0,2

0,674

0,04

0,00

0,01

BBa_808031

0,2

0,560

0,003

0

0,01

  • Midiprep of BBa_K808030
  • Spe I and PstI enzymatic digest of BBa_K808007 with the phosphatase step.
  • Gel migration on 1% agarose gel.
  • Gel Purification following the Macherey-Nagel Protocol
  • DNA concentration assay using a spectrophotometer.

Dilution 1/25

DNA Concentration (ng/µl)

OD(230nm)

OD(260nm)

OD(280nm)

OD(260nm)/OD(230nm)

OD(260nm)/OD(280nm

BBa_808007

0,8

0,07

0,017

0,014

0,24

1,25

BBa_808007

1,1

0,133

0,022

0,017

0,17

1,28

  • Ligation of BBa_K808007 and BBa_K808030
  • Transformation in electro-competent DH5-⍺ cells.
  • Preparation for sequencing.

Assembly of cluster 2 :

  • PCR amplification of BBa_K936011 and BBa_K936023 using Q5 High fidelity Master Mix.

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