Friday 12^th August

Lab work

Visualization

PCR of pPS16_009

By Léa

A Dreamtaq PCR was performed on pPS16_009 using the usual protocol. Primers: IPS83 ans IPS84

Electrophoresis of PCR products, using primers IPS 83 and IPS 84 on pPS16_016.

Extraction of pUC19

By Charlène

Clones 1 and 2 of pUC19 were extracted with the Plasmid MiniPrep kit.

They were fast digested with EcoRI : 3µL of plasmid + 1µL of FD buffer + 0.5µL of FD EcoRI + 5.5µL of water, 15 minutes at 37°C.

pUC19 clones 1 and 2 digested by EcoRI

Just clone 1 is at the good size so we will have to use it and not clone 2.

Dreamtaq PCR on puc19, detection, FRB, FKBP, SgRNAst, SgRNAnm and gBlock 1.2

By Naiane, Mahnaz and Terrence

A Dreamtaq PCR was performed on puc19, detection, FRB, FKBP, SgRNAst, SgRNAnm and gBlock 1.2 using the usual protocol with these volumes :

We made 6 clones of each colonie except for Puc19 where we made 1 negative control.

Result of the PCR : there is no band at the expected size for all the screened clones so the transformations of the gblock do not seem to have worked.

1.2 and FRB

Detection and ST

NM and FKBP

pub19 and FKBP