Difference between revisions of "Team:Paris Saclay/Notebook/July/28"
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= Thursday 28<sup>th</sup> July= | = Thursday 28<sup>th</sup> July= | ||
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===Visualization=== | ===Visualization=== | ||
| − | ====Low Fidelity DreamTaqPCR of | + | ====Low Fidelity DreamTaqPCR of DH5a|pPS16_001, DH5a|pPS16_002, DH5a|pPS16_005 and DH5a|pPS16_009==== |
| − | ''By Laetitia '' | + | ''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 [[Team:Paris_Saclay/Experiments#Polymerase_chain_reaction|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. | ||
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PCR was done with a Tm at 57°C. | PCR was done with a Tm at 57°C. | ||
| − | Each PCR | + | |
| + | Each PCR product was put on agarose gel to migrate. | ||
| + | 10µL of blue ladder was used. | ||
| + | |||
| + | PCR products expected were : | ||
| + | {| class="wikitable" | ||
| + | |- | ||
| + | !'''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. | ||
| + | |||
| + | |||
| + | [[File:T--Paris_Saclay--160728_visualization_PCR_Gblocks.jpeg|400px|thumb|right|Migration of pPS16_001 (Gblock 1.1) and pPS16_002 (Gblock 1.2)]] | ||
| + | [[File:T--Paris_Saclay--160728_visualization_PCR_Gblocks_GFP.jpeg|400px|thumb|right|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. | ||
| + | |||
| + | |||
| + | [[File:T--Paris_Saclay--160809_visualization3_-_Copie.jpeg|400px|thumb|right|Migration of Gblock 3.1 and GFP]] | ||
| + | [[File:T--Paris_Saclay--160809_visualization1.jpeg|400px|thumb|right|Migration of Gblock 1.1 and 1.2]] | ||
| + | [[File:T--Paris_Saclay--160809_visualization2.jpeg|200px|thumb|right|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. | ||
| + | |||
| + | {{Team:Paris_Saclay/notebook_footer}} | ||
Latest revision as of 16:11, 9 October 2016
