Team:Evry/Interlab Study/08-28-2014

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<h2>Contruction n°1: PSB1C3 with I20260</h2>
 
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<br/><h2> Contruction n°3: PSB1C3 with K823012-E1010</h2>
<br/><h2> Contruction n°3: PSB1C3 with K823012-E1010</h2>
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<br/>PCR colony was done on 7 colonies:
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<ol>
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<li>Steril H2O:34,5µL
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<li>5X Q5 Reaction buffer: 10µL
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<li>Template:2µL
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<li>10mM dNTP: 1µL
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<li>10µM Primer forward VF2: 1µL
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<li>10µM Primer reverse VR: 1µL
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<li> Q5 DNA Polymerase: 0,5µL
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</ol>
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<br/>A migration of PCR product was done. We expect to observe in each wells a band of 1176bp:
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<br/><img src="https://static.igem.org/mediawiki/2014/b/b6/Evry2014_J23115-E0240_gel_migration_.jpg" alt="image not found" />
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<br/> 6 colonies have a band at 1200 bp around so we decided to choose colonies n°2, 3 and 4.
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<h2>Other constructions of the Anderson library of constitutive promoters</h2>
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<h2>Constructions with other promoters</h2>
 
Promoters were resuspended from 2014 distribution kit plates with 10 µl of sterile MiliQ watter and stored at -20°C.
Promoters were resuspended from 2014 distribution kit plates with 10 µl of sterile MiliQ watter and stored at -20°C.
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<br/> Transformation in DH5 alpha chemocompetent <i>E. coli</i>.
 

Latest revision as of 02:04, 20 September 2014

Picture



Contruction n°2: PSB1C3 with J23101-E1010




Contruction n°3: PSB1C3 with K823012-E1010


PCR colony was done on 7 colonies:
  1. Steril H2O:34,5µL
  2. 5X Q5 Reaction buffer: 10µL
  3. Template:2µL
  4. 10mM dNTP: 1µL
  5. 10µM Primer forward VF2: 1µL
  6. 10µM Primer reverse VR: 1µL
  7. Q5 DNA Polymerase: 0,5µL

A migration of PCR product was done. We expect to observe in each wells a band of 1176bp:
image not found

6 colonies have a band at 1200 bp around so we decided to choose colonies n°2, 3 and 4.

Other constructions of the Anderson library of constitutive promoters


Promoters were resuspended from 2014 distribution kit plates with 10 µl of sterile MiliQ watter and stored at -20°C.
IMAGE
Symbol
Table: Additional promoters of the Interlab Study. In grey, plasmids already resuspended, transformed, sequenced and stock glycerol existed.

Aug 28

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