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

Line 104: Line 104:
 
                
 
                
 
<p style="text-align:left">1/ After 72h of incubation, only contamination was  observed.</p>
 
<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"> 2/ Co-transfection in yeast of nicked pRS415 nicked and Cre with extensions the protocol given by Heloise MÜLLER. </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>
+
<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>
 
<br>
 
<p style="text-align:left">Assembly of Cluster 1 :</p>
 
<p style="text-align:left">Assembly of Cluster 1 :</p>
 
<ul style="text-align:left">
 
<ul style="text-align:left">
−
   <li>Xba I and Pst I enzymatic digest. </li>
+
   <li>Xba I and Pst I enzymatic digestion. </li>
−
   <li>Gel migration on 1% agarose gel. </li>
+
   <li>Gel migration on 1% agarose gel. </li>
 
   <li>Gel Extraction following the  Macherey-Nagel Protocol </li>
 
   <li>Gel Extraction following the  Macherey-Nagel Protocol </li>
 
   <li>DNA Concentration </li>
 
   <li>DNA Concentration </li>
 
</ul>
 
</ul>
 +
</br>
 
<table border="1" cellspacing="0" cellpadding="0" width="604">
 
<table border="1" cellspacing="0" cellpadding="0" width="604">
 
   <tr>
 
   <tr>
−
     <td width="101" valign="top"><p>Dilution 1/25</p></td>
+
     <td width="101" valign="top"><p><b>Dilution 1/25</b></p></td>
−
     <td width="101" valign="top"><p>DNA Concentration (ng/µl)</p></td>
+
     <td width="101" valign="top"><p><b>DNA Concentration (ng/µl)</b></p></td>
−
     <td width="101" valign="top"><p>OD(230nm)</p></td>
+
     <td width="101" valign="top"><p><b>OD(230nm</b>)</p></td>
−
     <td width="101" valign="top"><p>OD(260nm)</p></td>
+
     <td width="101" valign="top"><p><b>OD(260nm)</b></p></td>
−
     <td width="101" valign="top"><p>OD(280nm)</p></td>
+
     <td width="101" valign="top"><p><b>OD(280nm)</b></p></td>
−
     <td width="101" valign="top"><p>OD(260nm)/OD(230nm)</p></td>
+
     <td width="101" valign="top"><p><b>OD(260nm)/OD(230nm)</b></p></td>
 
   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
−
     <td width="101" valign="top"><p>BBa_808030</p></td>
+
     <td width="101" valign="top"><p><b>BBa_K808030</b></p></td>
 
     <td width="101" valign="top"><p align="right">0,2 </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">0,674 </p></td>
Line 132: Line 133:
 
   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
−
     <td width="101" valign="top"><p>BBa_808031</p></td>
+
     <td width="101" valign="top"><p><b>BBa_K808031</b></p></td>
 
     <td width="101" valign="top"><p align="right">0,2 </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">0,560 </p></td>
Line 149: Line 150:
 
<table border="1" cellspacing="0" cellpadding="0" width="604">
 
<table border="1" cellspacing="0" cellpadding="0" width="604">
 
   <tr>
 
   <tr>
−
     <td width="86" valign="top"><p>Dilution 1/25</p></td>
+
     <td width="86" valign="top"><p><b>Dilution 1/25</b></p></td>
−
     <td width="86" valign="top"><p>DNA Concentration (ng/µl)</p></td>
+
     <td width="86" valign="top"><p><b>DNA Concentration (ng/µl)</b></p></td>
−
     <td width="86" valign="top"><p>OD(230nm)</p></td>
+
     <td width="86" valign="top"><p><b>OD (230 nm)</b></p></td>
−
     <td width="86" valign="top"><p>OD(260nm)</p></td>
+
     <td width="86" valign="top"><p><b>OD (260 nm)</b></p></td>
−
     <td width="86" valign="top"><p>OD(280nm)</p></td>
+
     <td width="86" valign="top"><p><b>OD (280 nm)</b></p></td>
−
     <td width="86" valign="top"><p>OD(260nm)/OD(230nm)</p></td>
+
     <td width="86" valign="top"><p><b>OD (260 nm) / OD (230 nm)</b></p></td>
−
     <td width="86" valign="top"><p>OD(260nm)/OD(280nm</p></td>
+
     <td width="86" valign="top"><p><b>OD (260 nm) / OD (280 nm)</b></p></td>
 
   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
−
     <td width="86" valign="top"><p>BBa_808007</p></td>
+
     <td width="86" valign="top"><p><b>BBa_K808007</b></p></td>
 
     <td width="86" valign="top"><p align="right">0,8 </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">0,07 </p></td>
Line 167: Line 168:
 
   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
−
     <td width="86" valign="top"><p>BBa_808007</p></td>
+
     <td width="86" valign="top"><p><b>BBa_K808007</b></p></td>
 
     <td width="86" valign="top"><p align="right">1,1 </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">0,133 </p></td>

Revision as of 17:25, 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 MÜLLER.

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 digestion.
  • 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_K808030

0,2

0,674

0,04

0,00

0,01

BBa_K808031

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 (230 nm)

OD (260 nm)

OD (280 nm)

OD (260 nm) / OD (230 nm)

OD (260 nm) / OD (280 nm)

BBa_K808007

0,8

0,07

0,017

0,014

0,24

1,25

BBa_K808007

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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