Difference between revisions of "Team:Paris Saclay/Notebook/July/28"

(Low Fidelity DreamTaqPCR of DH5a|pPS16_001, DH5a|pPS16_002, DH5a|pPS16_005 and DH5a|pPS16_009)
(Low Fidelity DreamTaqPCR of DH5a|pPS16_001, DH5a|pPS16_002, DH5a|pPS16_005 and DH5a|pPS16_009)
Line 42: Line 42:
  
  
[[File:T--Paris_Saclay--160728_visualization_PCR_Gblocks_GFP.jpeg|400px|thumb|right|Migration of pPS16_005 (Gblock 3.1) and pPS16_009 (GFP)]]
+
[File:T--Paris_Saclay--160809_visualization3_-_Copie.jpeg|400px|thumb|right|Migration of Gblock 3.1 and GFP ]]
  
 
===Biobrick Characterization===
 
===Biobrick Characterization===

Revision as of 13:12, 9 August 2016

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.


[File:T--Paris_Saclay--160809_visualization3_-_Copie.jpeg|400px|thumb|right|Migration of Gblock 3.1 and 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.