Wednesday 6^th July

Lab work

Preparation of LB solid and liquid stock

By Léa, Naiane, Laetitia

- 2L of LB liquid : 20g/L powder LB + 2L water μQ
- 1L of LB solid : 1L of LB liquid +15g/L of Agar
The all was put in the autoclave for sterilization with the help of Sylvain.

Biobrick characterization

DNA extraction of K1372001 clone 1 and clone 2

By Mathilde

Plasmids DNA were extracted following the usual protocol except that isopropanol was used instead of 100% ethanol and that pellets were dried on the lab bench top instead of in the speedvac. The extractions were kept at -20°C.

Visualization

gBlocks PCR screening

By Caroline

A PCR screening was carry out on the bactaria transformed with the gBlocks (6 clones for each except pPS16_004 for which it was only 2 clones). We use the Taq usual protocol. We used puc19 universal primers 1151_pheoR and 1152_pheoF.

1 μL of each culture were added for each Gblock pPS16_001, pPS16_002, pPS16_003, pPS16_004, pPS16_005, pPS16_006 and pPS16_007.

pPS16_004, pPS16_007 and the pPS16_007 clone 6 (selected on 01/07/2016) streaks

By Léa, Caroline and Laetitia

The transformations carried out the 30/06/2016 were streaked again on LB Petri dishes containing 50µg/ml of ampliciline on which X-Gal/IPTG diluted at 1/1000 was spread on (white/blue screen). The Petri dishes were incubated ON at 37°C.