Wednesday 24^th August

Lab work

Visualization

Gel Electrophoresis of Gibson (fragment 3 + fragment 4) and GFP1-9 cloned in PSB1C3

By Terrence

Clone 6 and 8 seems correct for the Gibson.

Clone 2-4-7-11 are correct.

pPS16_002 (1.2) clones 8 and 14 and pPS16_014(SgRna Nm) clone 12 Extraction

By Terrence

The extraction was carried out following the usual protocol.

Result of the extraction

Nano drop :

Transformation of pPS16_010 (FRB) in pJET and pPS16_014 (SgRnaNm) in pJET

By Terrence

First we made a ligation with

Then we transformed 2µL of ligation product into 50 µL of competent cells, following the usual protocol.

Transformation products were plated in duplicate : 50µL , 150µl and 350µL. 350µL only for the control.

Plasmid DNA extraction (Midi Prep)

By Léa & Manhaz

200 mL of transformed DH5a cells with Adgene dCas9 Nm plasmids (clone 1 and 2) cultures were extracted usind a Midi Prep kit. It was resuspended into 50µL of sterile water.

PCR Q5 on plasmid

By Léa & Manhaz