Team:Evry/Notebook/w7

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<h2>Wednesday, 2th July</h2>  
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<h2>Wednesday, 25th July</h2>  
Reception of primers fo Auxin Enzymes: IaaH FW and Rv and IaaM FW and RV.<br>
Reception of primers fo Auxin Enzymes: IaaH FW and Rv and IaaM FW and RV.<br>
PCR of imperial college BB with these primers to BB IaaH and IaaM.
PCR of imperial college BB with these primers to BB IaaH and IaaM.
Electrophoresis show that results aren't good.
Electrophoresis show that results aren't good.
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<h3>PCR:</h3>
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PCR of pCS2+:
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<table border="1">
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      <tr>
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        <td>Reactants</td>
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        <td>Volumes (µl)</td>
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      </tr>
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      <tr>
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        <td>GC Buffer</td>
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        <td>10</td>
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      </tr> 
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      <tr>
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        <td>dNTPs</td>
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        <td>1</td>
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      </tr> 
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      <tr>
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        <td>DNA</td>
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        <td>0,5</td>
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      </tr> 
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      <tr>
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        <td>H2O</td>
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        <td>33</td>
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      </tr> 
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      <tr>
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        <td>Primers (FW and RV)</td>
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        <td>2,5 each</td>
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      </tr> 
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      <tr>
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        <td>fusion polymerase</td>
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        <td>0,5</td>
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      </tr> 
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<table/>
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<h3>PCR:</h3>
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0,8% agarose
<h2>Thursday, 26th July</h2>
<h2>Thursday, 26th July</h2>

Latest revision as of 10:01, 9 August 2012

Weeks:

June July August September October November

Week 7: 23rd July - 29th July

Monday, 23rd July

Cloning:

4 different clones pCS2+ RFP => incubation in LB medium overnight at 37 degree celsius.

PCR:

PCR of pCS2+:
Reactants Volumes (µl)
GC Buffer 10
dNTPs 1
DNA 1
H2O 32,5
Primers (FW and RV) 2,5 each
fusion polymerase 0,5

Xenopus:

Start of auxin's toxicity test on tadpoles: Tadpoles in there growth media + 0/125/250 or 500 µM auxin
We changed this media each days during one week

Tuesday, 24th July

Plasmid Purification:

On the pCS2+ RFP clones incubated the day before:
DNA Concentration
Tube
Concentration (ng.uL-1)
1
144,2
2
161,9
3
184,9
4
120,7

Digestion:

Plasmid digestion
Tube
V DNA (uL)
V SpeI (uL)
V EcoRI (uL)
V buffer (uL)
V H2O (uL)
1
7
1
1
2
9
2
6,2
1
1
2
9,8
3
5,4
1
1
2
10,6
4
8,3
1
1
2
7,7

Gel migration:

Gel 0,8%

Wednesday, 25th July

Reception of primers fo Auxin Enzymes: IaaH FW and Rv and IaaM FW and RV.
PCR of imperial college BB with these primers to BB IaaH and IaaM. Electrophoresis show that results aren't good.

PCR:

PCR of pCS2+:
Reactants Volumes (µl)
GC Buffer 10
dNTPs 1
DNA 0,5
H2O 33
Primers (FW and RV) 2,5 each
fusion polymerase 0,5

PCR:

0,8% agarose

Thursday, 26th July

Test of auxin toxicity in tadpodes. Retry of IaaH and IaaM BB: Ok for IaaM but not for IaaH.

Friday, 27th July

  • PCR of TirI: One PCR with primers TirI FW + Sdm RV / One PCR with primers TirI RV + Sdm FW
  • Gel extraction of these PCR
  • New PCR with the mix of the two PCR products + primers TirI FW and RV