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 Gblock 3.1 and GFP

Migration of Gblock 1.1 and 1.2

Migration of Gblock 1.1 and 1.2

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.