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 8th August 1.1 Visualization 1.1.1 Phusion PCR on pPS16_006 and pPS16_007 1.1.2 Liquid cultures of bacteria containing plasmid to extract 1.1.3 Migration of DNA plasmid extracted Monday 8th August Visualization Phusion PCR on pPS16_006 and pPS16_007 By Caroline The PCR was carried out following the usual protocol adapted to 50µL and with a TM at 72°C. The specific primers for each parts were used. The products were put for migration on a 0.8%agarose gel with BET. No amplification were observed: that was probably due to the fact that the plasmid extractions made on the 3/08/2016 were eluted with water which is not working with this plasmid extraction kit. Result of the PCR Liquid cultures of bacteria containing plasmid to extract By Caroline, Charlène, Terrence In order to redo the plasmid extractions that did not work due to the use of water instead of the elution buffer. To do it, 5mL of LB were mixed with Ampicillin at 50µg/mL and put at 37°C and at 180rpm overnight. Migration of DNA plasmid extracted By Terrence, Alice, Laetitia Plasmid sent to sequencing were migrated again on a gel in order to check their size. A lot of depositions have no DNA which would be explained by the use of water instead of the elution buffer of the plasmid extraction kit. Migration of pPS16_001 (Gblock 1.1), pPS16_002 (Gblock 1.2), pPS16_005, pPS16_006, pUC 19 Migration
By Caroline
The PCR was carried out following the usual protocol adapted to 50µL and with a TM at 72°C. The specific primers for each parts were used. The products were put for migration on a 0.8%agarose gel with BET. No amplification were observed: that was probably due to the fact that the plasmid extractions made on the 3/08/2016 were eluted with water which is not working with this plasmid extraction kit.
By Caroline, Charlène, Terrence
In order to redo the plasmid extractions that did not work due to the use of water instead of the elution buffer. To do it, 5mL of LB were mixed with Ampicillin at 50µg/mL and put at 37°C and at 180rpm overnight.
By Terrence, Alice, Laetitia
Plasmid sent to sequencing were migrated again on a gel in order to check their size. A lot of depositions have no DNA which would be explained by the use of water instead of the elution buffer of the plasmid extraction kit.
