Team:Evry/Notebook/w7

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<h1>Weeks</h1>
 
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<table>
 
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<tr>
 
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  <td><a href="https://2012.igem.org/Team:Evry/Notebook/w1">1</a></td>
 
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  <td><a href="https://2012.igem.org/Team:Evry/Notebook/w2">2</a></td>
 
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  <td><a href="https://2012.igem.org/Team:Evry/Notebook/w3">3</a></td>
 
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  <td><a href="https://2012.igem.org/Team:Evry/Notebook/w4">4</a></td>
 
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  <td><a href="https://2012.igem.org/Team:Evry/Notebook/w5">5</a></td>
 
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  <td><a href="https://2012.igem.org/Team:Evry/Notebook/w6">6</a></td>
 
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  <td>7</td>
 
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  <td><a href="https://2012.igem.org/Team:Evry/Notebook/w8">8</a></td>
 
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  <td><a href="https://2012.igem.org/Team:Evry/Notebook/w9">9</a></td>
 
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  <td><a href="https://2012.igem.org/Team:Evry/Notebook/w10">10</a></td>
 
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  <td><a href="https://2012.igem.org/Team:Evry/Notebook/w11">11</a></td>
 
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  <td><a href="https://2012.igem.org/Team:Evry/Notebook/w12">12</a></td>
 
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  <td><a href="https://2012.igem.org/Team:Evry/Notebook/w13">13</a></td>
 
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  <td><a href="https://2012.igem.org/Team:Evry/Notebook/w14">14</a></td>
 
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  <td><a href="https://2012.igem.org/Team:Evry/Notebook/w15">15</a></td>
 
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  <td><a href="https://2012.igem.org/Team:Evry/Notebook/w16">16</a></td>
 
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  <td><a href="https://2012.igem.org/Team:Evry/Notebook/w17">17</a></td>
 
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  <td><a href="https://2012.igem.org/Team:Evry/Notebook/w18">18</a></td>
 
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  <td><a href="https://2012.igem.org/Team:Evry/Notebook/w19">19</a></td>
 
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  <td><a href="https://2012.igem.org/Team:Evry/Notebook/w20">20</a></td>
 
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  <td><a href="https://2012.igem.org/Team:Evry/Notebook/w21">21</a></td>
 
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  <td><a href="https://2012.igem.org/Team:Evry/Notebook/w22">22</a></td>
 
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</table>
 
<h1>Week 7: 23rd July - 29th July</h1>
<h1>Week 7: 23rd July - 29th July</h1>
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<h3>Gel migration:</h3>
<h3>Gel migration:</h3>
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Gel 0,8%  
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Gel 0,8%
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<h2>Wednesday, 25th July</h2>
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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.
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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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        <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>
Test of auxin toxicity in tadpodes.
Test of auxin toxicity in tadpodes.
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Retry of IaaH and IaaM BB: Ok for IaaM but not for IaaH.
<h2>Friday, 27th July</h2>
<h2>Friday, 27th 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

Retrieved from "http://2012.igem.org/Team:Evry/Notebook/w7"