Monday 8^th August

Lab work

Visualization

Phusion PCR on pPS16_006 and pPS16_007

By Caroline

The PCR was carried out following the usual protocol adapted to 50µL and with a TM at 72°C. The specific primers for each parts were used. The products were put for migration on a 0.8%agarose gel with BET. No amplification were observed: that was probably due to the fact that the plasmid extractions made on the 3/08/2016 were eluted with water which is not working with this plasmid extraction kit.

Result of the PCR

Liquid cultures of bacteria containing plasmid to extract

By Caroline, Charlène, Terrence

In order to redo the plasmid extractions that did not work due to the use of water instead of the elution buffer. To do it, 5mL of LB were mixed with Ampicillin at 50µg/mL and put at 37°C and at 180rpm overnight.

Migration of DNA plasmid extracted

By Terrence, Alice, Laetitia

Plasmid sent to sequencing were migrated again on a gel in order to check their size. A lot of wells have no DNA which would be explained by the use of water instead of the elution buffer of the plasmid extraction kit.

Migration of pPS16_001 (Gblock 1.1), pPS16_002 (Gblock 1.2), pPS16_005, pPS16_006, pUC 19

Migration