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 placed on agarose gel to migrate. 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) Migration of pPS16_005 (Gblock 3.1) and pPS16_009 (GFP) 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. 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 placed on agarose gel to migrate. 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.
