Thursday 28^th 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 :

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.

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.