Wednesday 3^rd August

Lab work

Visualization

Glycerol stocks

By Caroline

The bacteria transformed with the plasmids sent the 2/08/2016 and those that would be sent the 4/08/2016 were put into glycerol. 1mL of liquid culture and 0.5mL of glycerol were put at -20°C.

High fidelity Q5 PCR on pPS16_004, pPS16_006 and pPS16_007

By Caroline

Q5 PCR was carried out following the usual protocol adapted to 50µL at a Tm of 72°C. The primers used were specific to amplify only the sequences of interest. The products were put for migration on a 0.8%agarose gel with BET. Only, PCR form pPS16_004 and pPS16_007 showed a positive result.

Extraction of the plasmids containing gBlocks pPS16_001, pPS16_002, pPS16_003, FRB, sg-ST1, FKPB, DS-NMcasN-, spacer and detection

By Laetitia

The extraction was performed without kit, following the usual protocol.

It was done on:

Extraction of the plasmids containing gBlocks

By Naiane

The kit "Charge Switch-Pro Plasmid Miniprep" was used to extract plasmidic DNA of the plasmid pPS16_003 containing the Gblock 2.1, plasmid pPS16_004 containing the Gblock 2.2, plasmid pPS16_006 containing the Gblock 3.2 and the plasmid pPS16_007 containing the Gblock 4.1 from 3mL of overnight culture. Plasmids were resuspended in 100μL of Milli-Q water. DNA stored at -20°C.

Interlab Study

Preculture of device one transformed DH5a

By Lea

Two colonies were inoculated inside of two tubes of 3mL of liquid LB medium containing 30 µg/ml Chloramphenicol. The tubes were incubated at 37°C overnight.