Toggle navigation Home Team Team Media Collaborations Sponsors Acknowledgements Project Background Design CRISPR/Cas9 Strategy Experiments Notebook Results Perspective Interlab Study Parts Parts Basic Parts Composite Parts Human Pratices Overview Societal Issues of CRISPR/Cas9 Responsible Research and Innovation GMO regulation Integrated Practices Engagement Model Attributions Safety Contents 1 Friday 12th August 1.1 Lab work 1.1.1 Visualization 1.1.1.1 PCR of pPS16_009 1.1.1.2 Extraction of puc19 1.1.1.3 Dreamtaq PCR on puc19, detection, FRB, FKBP, SgRNAst, SgRNAnm and gBlock 1.2 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.
By Léa
A Dreamtaq PCR was performed on pPS16_009 using the usual protocol. Primers: IPS83 ans IPS84
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.
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.
