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− No PCR products are observed, due to an error in the protocol : MilliQ water was used instead of steril water.
+ 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 .
Revision as of 15:24, 12 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.
