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 Friday 5st August 1.1 Lab work 1.1.1 Visualization 1.1.1.1 DNA Extraction of DS-TDcasN- and DS-SPcasN- 1.1.1.2 DNA Extraction of DS-NMcasN- and pPS_001 1.1.1.3 Q5 PCR on pPS16_003, pPS16_004, pPS16_006 and pPS16_007 1.1.1.4 PCR Clean-up with the NucleoSpin kit Friday 5st August Lab work Visualization DNA Extraction of DS-TDcasN- and DS-SPcasN- By Laetitia The extraction was performed using the kit. We followed this protocol. The initial culture was at 15 mL so we increased (*4) the volumes until the precipitation. Hence we used: -1 ml of resuspension buffer -1 ml lysis buffer -1 ml precipitation buffer The colum was used several times in order to recover the maximum of DNA DNA Extraction of DS-NMcasN- and pPS_001 By Mathilde The extraction was performed on clone 11 for NM and clone 9 for 1.1 using the kit. We followed this protocol. Q5 PCR on pPS16_003, pPS16_004, pPS16_006 and pPS16_007 By Caroline The PCR was carried out following the usual protocol adapted to 50µL and with a TM at 70°C. The specific primers for each parts were using. The products were put to migrated on a 0.8%agarose gel with BET. Only, PCR form pPS16_003 and pPS16_004 showed a positive result. PCR Clean-up with the NucleoSpin kit By Caroline The purification was carried out on DH5alp|pPS16_003, DH5alp|pPS16_004 and DH5alp|pPS16_007 following the usual protocol. But, I forgot to diluted the Buffet NT3 with 100mL ethanol befor I began the purification and so any result was obtained on the 0.8% agarose gel.
By Laetitia
The extraction was performed using the kit. We followed this protocol.
The initial culture was at 15 mL so we increased (*4) the volumes until the precipitation.
Hence we used:
-1 ml of resuspension buffer
-1 ml lysis buffer
-1 ml precipitation buffer
The colum was used several times in order to recover the maximum of DNA
By Mathilde
The extraction was performed on clone 11 for NM and clone 9 for 1.1 using the kit. We followed this protocol.
By Caroline
The PCR was carried out following the usual protocol adapted to 50µL and with a TM at 70°C. The specific primers for each parts were using. The products were put to migrated on a 0.8%agarose gel with BET. Only, PCR form pPS16_003 and pPS16_004 showed a positive result.
The purification was carried out on DH5alp|pPS16_003, DH5alp|pPS16_004 and DH5alp|pPS16_007 following the usual protocol. But, I forgot to diluted the Buffet NT3 with 100mL ethanol befor I began the purification and so any result was obtained on the 0.8% agarose gel.
