Line 10:
Line 10: Primers: IPS83 ans IPS84
Primers: IPS83 ans IPS84
− [[File:T--Paris_Saclay--120826_visualization_PCRGFP.jpeg|400px|thumb|right|Electrophoresis of PCR products, using primers IPS 83 and IPS 84 on pPS16_016.]]
+ [[File:T--Paris_Saclay--120826_visualization_PCRGFP.jpg |400px|thumb|right|Electrophoresis of PCR products, using primers IPS 83 and IPS 84 on pPS16_016.]]
====Extraction of puc19====
====Extraction of puc19====
Revision as of 14:29, 16 August 2016
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
Electrophoresis of PCR products, using primers IPS 83 and IPS 84 on pPS16_016.
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.
