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 Monday 4th July 1.1 Lab work 1.1.1 Visualization 1.1.1.1 Digestion of plasmids 1.1.2 Bringing DNA closer 1.1.2.1 Plasmids extraction Monday 4th July Lab work Visualization Digestion of plasmids By Mathilde and Alice The following plasmids were digested again with EcoR1 and HindIII: 1.1 (from the 6 clones that were selected on 29/06/16) 2.1 (from the 6 clones that were selected on 29/06/16) 3.1 (from the 6 clones that were selected on 29/06/16) 3.2 (from the 6 clones that were selected on 29/06/16) Component Volume (µL) Plasmid 10 Red buffer 10x 2 Water 7 EcoRI enzyme 0.5 HindIII enzyme 0.5 The mix was incubated at 37°C for 1 hour. An electrophoresis was done (0.8% of agarose). Component Volume (µL) Digested DNA 20 Loading buffer 3.3 The DNA concentration was still insufficient meaning the extraction step was not efficient. Colony PCR Bringing DNA closer Plasmids extraction By Naiane and Laetitia Plasmids from the following strains were extracted with the kit "Charge Switch-Pro Plasmid Miniprep": DS-NMCas DS-SPCasN DS-ST1 DS-TDCasN Then plasmids were digested with AvrII increasing the DNA quantity (10µL DNA, 1µL AvrII, 2µL Tango buffer, 7µL water). Digestion products were migrated (5µL digestion, 5µL water, 2µL Template:Font color
By Mathilde and Alice
The following plasmids were digested again with EcoR1 and HindIII:
The mix was incubated at 37°C for 1 hour. An electrophoresis was done (0.8% of agarose).
The DNA concentration was still insufficient meaning the extraction step was not efficient. Colony PCR
By Naiane and Laetitia
Plasmids from the following strains were extracted with the kit "Charge Switch-Pro Plasmid Miniprep":
Then plasmids were digested with AvrII increasing the DNA quantity (10µL DNA, 1µL AvrII, 2µL Tango buffer, 7µL water). Digestion products were migrated (5µL digestion, 5µL water, 2µL Template:Font color
