Tuesday 23^th August

Lab work

Visualization

Colony PCR

By Léa and Naiane

12 colonies containing the gBlock GFP 1-9 and 12 colonies transformed with Gibson Assembly products (fragment 3-4) were screened and used for the usual protocol of Colony PCR. The same colonies were also plated on Petri dish containing solid LB + Cm and liquid culture (LB + Cm) was made from these same colonies. Both petri dishes and the liquid cultures were overnight at 37°C. The migration on agarose gel was done the next day.

Cell Cultures

By Léa, Naiane and Mathilde

100µL ofDH5a precultures containing dCas9 Nm plasmids (clone 1 and 2) were used to inoculate 200mL of liquid LB medium containing spectinomycin.

Interlab study plates from the 22/08/2016 were used to inoculate two tubes containg 5mL of liquid LB medium (chloramphenicol) per construction.

DH5a colonies containing pUC19 vector cloned with sgRNA NM (clone 5 and 12) or 1.2 (clones 8 and 14) were inoculated into 4mL of liquid LB medium.