Team:Evry/Notebook/w8

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<h1>Weeks</h1>
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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/w7">7</a></td>
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  <td>8</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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<h1>Week 8: 30th July - 5th August</h1>
<h1>Week 8: 30th July - 5th August</h1>
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Ligation and Transformation on T10.  
Ligation and Transformation on T10.  
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<h2>Tuesday, 31th July</h2>
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<h3>TirI BB</h3>
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Electrophoresis of TirI last PCR: failure of this technique.<br>
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We buy a new primer to try another PCR technic.
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<h2>Tuesday, 31st July</h2>
<h3>Inoculation:</h3>
<h3>Inoculation:</h3>
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Electrophoresis of 27/07 results: Failure of second PCR</br>
Electrophoresis of 27/07 results: Failure of second PCR</br>
We restart the first step  
We restart the first step  
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<h2>Thursday, 2nd August</h2>
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<h3>Sequencing</h3>
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Send pCS2+ to sequencing.
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<h3>Issue</h3>
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We found that pCS2+ was badly made. We restart the construction of this plasmid from the beginning.
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<h2>Friday, 3nd August</h2>
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<h3>Construction pCS2+ biobricked:</h3>
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PCR pCS2+ Biobricked
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Result gel:<br/>
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1) is PCR products with template pCS2+, primers P7 and P8    and  2) to 6) are ladders test
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<br/><br/>
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Then the PCR product pCS2+ was purified with kit, digested with EcoRI and PstI then purified with PCR cleanup kit.<br/>
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Plasmid TSO 28.06 mRFP (plasmid from plate 1-18F, mRFP)was cut with EcoRI and PstI:
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after electrophoresis no band visible, 7 microL of plasmid used in a total volume of 25 microL.
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Latest revision as of 18:19, 3 August 2012

Weeks:

June July August September October November

Week 8: 30th July - 5th August

Monday, 30th July

Digestion - Ligation - Transformation:

Digestion of psc2+ and mRFP with EcoRI and PstI.
Ligation and Transformation on T10.

TirI BB

Electrophoresis of TirI last PCR: failure of this technique.
We buy a new primer to try another PCR technic.

Tuesday, 31st July

Inoculation:

Inoculation of pcs2+ (27.07) and repiquage of psc2+ with RFP (30.07)
Inoculation of psB1C3 with GFP-AID TirI: Electrophoresis of 27/07 results: Failure of second PCR
We restart the first step

Thursday, 2nd August

Sequencing

Send pCS2+ to sequencing.

Issue

We found that pCS2+ was badly made. We restart the construction of this plasmid from the beginning.

Friday, 3nd August

Construction pCS2+ biobricked:

PCR pCS2+ Biobricked Result gel:
A gel width=
1) is PCR products with template pCS2+, primers P7 and P8 and 2) to 6) are ladders test

Then the PCR product pCS2+ was purified with kit, digested with EcoRI and PstI then purified with PCR cleanup kit.
Plasmid TSO 28.06 mRFP (plasmid from plate 1-18F, mRFP)was cut with EcoRI and PstI: after electrophoresis no band visible, 7 microL of plasmid used in a total volume of 25 microL.