Team:Paris Saclay/Notebook/July/28

Thursday 28th July

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 spread on a Petri dish (LB+AMP+X-Gal+IPTG.

PCR was done with a Tm at 57°C.

Each PCR product was put on agarose gel to migrate. 10µL of blue ladder was used.

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 spread 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.