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 Tuesday 3st August 1.1 Lab work 1.1.1 Visualization 1.1.1.1 Glycerol stocks 1.1.1.2 High fidelity Q5 PCR on DH5alp|pPS16_004, DH5alp|pPS16_006 and DH5alp|pPS16_007 1.1.1.3 1.1.1.4 Plasmid extraction 1.1.2 Interlab Study 1.1.2.1 Tuesday 3st August Lab work Visualization Glycerol stocks By Caroline The bacteria transformed with the plasmids sent the 2/08/2016 and those that would be sent the 4/08/2016 were put into glycerol. 1mL of liquid culture and 0.5mL of glycerol were put at -20°C. High fidelity Q5 PCR on DH5alp|pPS16_004, DH5alp|pPS16_006 and DH5alp|pPS16_007 By Caroline Q5 PCR was carried out following the usual protocol adapted to 50µL at a TM of 72°C. The primers used were specific to amplify only the interested sequence. By Plasmid extraction By Naiane The usual protocol was used to extract plasmidic DNA of pPS16_003, pPS16_004, pPS16_006 and pPS16_007 (Gblocks 2.1, 2.2, 3.2 and 4.1) from 3mL of overnight culture. Plasmids were resuspended in 100μL of Milli-Q water. Interlab Study By
By Caroline
The bacteria transformed with the plasmids sent the 2/08/2016 and those that would be sent the 4/08/2016 were put into glycerol. 1mL of liquid culture and 0.5mL of glycerol were put at -20°C.
Q5 PCR was carried out following the usual protocol adapted to 50µL at a TM of 72°C. The primers used were specific to amplify only the interested sequence.
By
By Naiane
The usual protocol was used to extract plasmidic DNA of pPS16_003, pPS16_004, pPS16_006 and pPS16_007 (Gblocks 2.1, 2.2, 3.2 and 4.1) from 3mL of overnight culture. Plasmids were resuspended in 100μL of Milli-Q water.
