Difference between revisions of "Team:Paris Saclay/Notebook/July/8"

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No PCR products are observed, due to an error in the protocol : MilliQ water was used instead of steril water. The same PCR will be performed on 11/07/16.
 
No PCR products are observed, due to an error in the protocol : MilliQ water was used instead of steril water. The same PCR will be performed on 11/07/16.
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====Electrophoresis of the screening PCR products====
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''By Caroline''
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Amplification products from the 07/07/2016 screening PCR were put in a 0.8% agarose gel containing BET. The products in solution were mixed with purple loading dye and migrated for 30min.
 +
Some clones showed PCR products with lengths close to those expected.

Revision as of 08:09, 13 July 2016

Tuesday 8th July

Lab work

Visualization

Colony screening PCR
on bacteria transformed with pPS16_004 and pPS16_007

By Alice

After transformation, only white bacteria are selected (blue white screen). They are expected to have plasmids with inserts of interest. PCR with DreamTaq DNA Polymerase is performed following this protocol. Clones selected for PCR are kept on LB+Amp+XGal(2µLXGal/1000µLH2O)+IPTG(2µLIPTG/1000µLH2O). Two primers (1152_pheoF and 1151 5_pheoR) upstream and downstream the insertion site are chosen. . PCR products expected below :

Plasmids Band size (bp)
pPS16_004 846
pPS16_007 744

No PCR products are observed, due to an error in the protocol : MilliQ water was used instead of steril water. The same PCR will be performed on 11/07/16.

Electrophoresis of the screening PCR products

By Caroline

Amplification products from the 07/07/2016 screening PCR were put in a 0.8% agarose gel containing BET. The products in solution were mixed with purple loading dye and migrated for 30min. Some clones showed PCR products with lengths close to those expected.