Team:Paris Saclay/Notebook/August/12

Friday 12th 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

File:T--Paris Saclay--120826 visualization PCRGFP.jpeg
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.


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 :

Components Volume
10X DreamTaq Green Buffer 2.5µL
dNTP (10mM) 1µL
Primers mix (10µM each) 1µL
DreamTaq DNA polymerase 0.25µL
Nuclease-free water up to 25µL
Total volume 25µL

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