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 Wednesday 6th July 1.1 Lab work 1.1.1 Preparation of LB solid and liquid stock 1.1.2 Biobrick characterization 1.1.2.1 DNA extraction of K1372001 clone 1 and clone 2 1.1.3 Visualization 1.1.3.1 gBlocks PCR screening 1.1.3.2 pPS16_004, pPS16_007 and the pPS16_007 clone 6 (selected on 01/07/2016) streaks Wednesday 6th July Lab work Preparation of LB solid and liquid stock By Léa, Naiane, Laetitia - 2L of LB liquid : 20g/L powder LB + 2L water μQ - 1L of LB solid : 1L of LB liquid +15g/L of Agar The all was put in the autoclave for sterilization with the help of Sylvain. Biobrick characterization DNA extraction of K1372001 clone 1 and clone 2 By Mathilde Plasmids DNA were extracted following the usual protocol except that isopropanol was used instead of 100% ethanol and that pellets were dried on the lab bench top instead of in the speedvac. The extractions were kept at -20°C. Visualization gBlocks PCR screening By Caroline A PCR screening was carry out on the bactaria transformed with the gBlocks (6 clones for each except pPS16_004 for which it was only 2 clones). We use the Taq usual protocol. We used puc19 universal primers 1151_pheoR and 1152_pheoF. 1 μL of each culture were added for each Gblock pPS16_001, pPS16_002, pPS16_003, pPS16_004, pPS16_005, pPS16_006 and pPS16_007. pPS16_004, pPS16_007 and the pPS16_007 clone 6 (selected on 01/07/2016) streaks By Léa, Caroline and Laetitia The transformations carried out the 30/06/2016 were streaked again on LB Petri dishes containing 50µg/ml of ampliciline on which X-Gal/IPTG diluted at 1/1000 was spread on (white/blue screen). The Petri dishes were incubated ON at 37°C.
By Léa, Naiane, Laetitia
- 2L of LB liquid : 20g/L powder LB + 2L water μQ - 1L of LB solid : 1L of LB liquid +15g/L of Agar The all was put in the autoclave for sterilization with the help of Sylvain.
By Mathilde
Plasmids DNA were extracted following the usual protocol except that isopropanol was used instead of 100% ethanol and that pellets were dried on the lab bench top instead of in the speedvac. The extractions were kept at -20°C.
By Caroline
A PCR screening was carry out on the bactaria transformed with the gBlocks (6 clones for each except pPS16_004 for which it was only 2 clones). We use the Taq usual protocol. We used puc19 universal primers 1151_pheoR and 1152_pheoF.
1 μL of each culture were added for each Gblock pPS16_001, pPS16_002, pPS16_003, pPS16_004, pPS16_005, pPS16_006 and pPS16_007.
By Léa, Caroline and Laetitia
The transformations carried out the 30/06/2016 were streaked again on LB Petri dishes containing 50µg/ml of ampliciline on which X-Gal/IPTG diluted at 1/1000 was spread on (white/blue screen). The Petri dishes were incubated ON at 37°C.
