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) The same digestion protocol as on Template:Font color was followed increasing the DNA quantity (10µL DNA, 7µL water, 2µL Red buffer, 0.5µL EcoR1, 0.5µL HindIII). Digestion products were migrated on agarose gel. The DNA concentration was still insufficient meaning the extraction step was not efficient. New extraction will be performed on clones got after the transformation. 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 same digestion protocol as on Template:Font color was followed increasing the DNA quantity (10µL DNA, 7µL water, 2µL Red buffer, 0.5µL EcoR1, 0.5µL HindIII). Digestion products were migrated on agarose gel. The DNA concentration was still insufficient meaning the extraction step was not efficient. New extraction will be performed on clones got after the transformation.
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
