Wednesday 24^th August

Lab work

Visualization

Gel Electrophoresis of Gibson Assembly(fragment 3-4) and GFP 1-9 cloned in PSB1C3

By Terrence

Colony PCR of 12 colonies containing the Gibson Assembly's products (fragment 3-4).

Colony PCR of 12 colonies containing the GFP 1-9.

- Clones 2, 4, 7 and 11 are as expected.

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 gblockFRB in pJET and gblock Sg_RNA_NM in pJET

By Terrence

First we made a ligation with

Then we transformed 2µL of ligation product into 50 µL of DH5a competent cells, following the usual protocol.

Transformation products were plated with three different volume : 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