Difference between revisions of "Team:Paris Saclay/Notebook/August/10"
MahnaziGEM (Talk | contribs) (→Colony screening PCR on bacteria transformed with pPS16_010 and pPS16_011) |
MahnaziGEM (Talk | contribs) (→Colony screening PCR on bacteria transformed with pPS16_010 and pPS16_011) |
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====Colony screening PCR on bacteria transformed with pPS16_010 and pPS16_011==== | ====Colony screening PCR on bacteria transformed with pPS16_010 and pPS16_011==== | ||
''By Alice'' | ''By Alice'' | ||
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After transformation, only white bacteria were selected (blue and white screen). | After transformation, only white bacteria were selected (blue and white screen). | ||
For bacteria transformed with pPS_011, the only clone white was chosen, and for bacteria transformed with pPS16_010, 9 white clones among others were chosen. They are expected to have plasmids with the wanted inserts. | For bacteria transformed with pPS_011, the only clone white was chosen, and for bacteria transformed with pPS16_010, 9 white clones among others were chosen. They are expected to have plasmids with the wanted inserts. | ||
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PCR with DreamTaq DNA Polymerase was performed following [[Team:Paris_Saclay/Experiments#taqPCR|this protocol]]. Clones selected for PCR were kept on LB+Amp+XGal(2µLXGal/1000µLH2O)+IPTG(2µLIPTG/1000µLH2O). Two primers ([[Team:Paris_Saclay/Experiments#primers|1151_pheoR and 1152_pheoF]]) upstream and downstream the insertion site were chosen. Annealing temperature was 53°C. After amplification, 1 µL of each PCR products and 10 µL of DNA ladder were placed in wells and migrated at 100v during 30 min. This gel was also used to migrate pUC19 plasmids digested with HincII.<div id="August_10"></div> | PCR with DreamTaq DNA Polymerase was performed following [[Team:Paris_Saclay/Experiments#taqPCR|this protocol]]. Clones selected for PCR were kept on LB+Amp+XGal(2µLXGal/1000µLH2O)+IPTG(2µLIPTG/1000µLH2O). Two primers ([[Team:Paris_Saclay/Experiments#primers|1151_pheoR and 1152_pheoF]]) upstream and downstream the insertion site were chosen. Annealing temperature was 53°C. After amplification, 1 µL of each PCR products and 10 µL of DNA ladder were placed in wells and migrated at 100v during 30 min. This gel was also used to migrate pUC19 plasmids digested with HincII.<div id="August_10"></div> | ||
Revision as of 13:47, 17 August 2016
