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. 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.