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 Thursday 28th July 1.1 Lab work 1.1.1 Visualization 1.1.1.1 Low Fidelity DreamTaqPCR of DH5a|pPS16_001, DH5a|pPS16_002, DH5a|pPS16_005 and DH5a|pPS16_009 1.1.1.2 Low Fidelity DreamTaqPCR of DH5a|pPS16_001, DH5a|pPS16_002, DH5a|pPS16_005 and DH5a|pPS16_009 1.1.2 Biobrick Characterization 1.1.2.1 BL21 electrocompetent cells in glycerol stock Thursday 28th July Lab work Visualization Low Fidelity DreamTaqPCR of DH5a|pPS16_001, DH5a|pPS16_002, DH5a|pPS16_005 and DH5a|pPS16_009 By Mathilde and Laetitia PCR was performed on 6 clones for each plasmid. Thus, the PCR mix was done for 24 tubes following the usual protocol. Each clone was taken off from the petri dish (27/07) soaked in a PCR mix and finally re-plated on a Petri dish (LB+AMP+X-Gal+IPTG. PCR was done with a Tm at 57°C. Each PCR Product was put to migrate on an agarose gel. 10µL of blue ladder was used, and the wells were filled with 1µL of PCR preparations. PCR products expected were : Plasmid pPS16_001 pPS16_002 Band Size (bp) 1007 1007 Only pPS16_001 clone 3 and pPS16_002 clone 1 have amplified at the expected size. Migration of pPS16_001 (Gblock 1.1) and pPS16_002 (Gblock 1.2) Low Fidelity DreamTaqPCR of DH5a|pPS16_001, DH5a|pPS16_002, DH5a|pPS16_005 and DH5a|pPS16_009 By Mathilde, Laetitia and Caroline The exact same experiment as previously in the day was made, but with 6 different clones for each cultures. Each clone was taken off from the petri dish (27/07) soaked in a PCR mix and finally re-plated on a Petri dish (LB+AMP+X-Gal+IPTG. Each clone was also put in culture in 1,5mL of LB and put in incubation at 37°c, 180 rpm ON. Migration of pPS16_005 (Gblock 3.1) and pPS16_009 (GFP) Biobrick Characterization BL21 electrocompetent cells in glycerol stock By Charlène 1 mL of each clone cultured yesterday were put in 500µL of glycerol 60%. They were conserved at -30°C.
By Mathilde and Laetitia
PCR was performed on 6 clones for each plasmid.
Thus, the PCR mix was done for 24 tubes following the usual protocol.
Each clone was taken off from the petri dish (27/07) soaked in a PCR mix and finally re-plated on a Petri dish (LB+AMP+X-Gal+IPTG.
PCR was done with a Tm at 57°C.
Each PCR Product was put to migrate on an agarose gel. 10µL of blue ladder was used, and the wells were filled with 1µL of PCR preparations.
PCR products expected were :
Only pPS16_001 clone 3 and pPS16_002 clone 1 have amplified at the expected size.
By Mathilde, Laetitia and Caroline
The exact same experiment as previously in the day was made, but with 6 different clones for each cultures. Each clone was taken off from the petri dish (27/07) soaked in a PCR mix and finally re-plated on a Petri dish (LB+AMP+X-Gal+IPTG. Each clone was also put in culture in 1,5mL of LB and put in incubation at 37°c, 180 rpm ON.
By Charlène
1 mL of each clone cultured yesterday were put in 500µL of glycerol 60%. They were conserved at -30°C.
