Difference between revisions of "Team:Paris Saclay/Notebook/July/22"
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=Friday 22<sup>nd</sup> July= | =Friday 22<sup>nd</sup> July= | ||
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| − | === | + | ===Visualization=== |
| − | + | ====Low Fidelity Dreamtaq PCR of DH5α|pPS16_002==== | |
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| − | ====Low Fidelity Dreamtaq PCR of | + | |
''By Mathilde'' | ''By Mathilde'' | ||
| − | A DreamTaq PCR was made with [[Team:Paris_Saclay/Notebook/July/19#Visualization| | + | A DreamTaq PCR was made with [[Team:Paris_Saclay/Notebook/July/19#Visualization|DH5α|pPS16_002]] cultures following the usual [[Team:Paris_Saclay/Experiments#Polymerase_chain_reaction|protocol]] with Tm at 57°C. |
| − | We divided up the PCR mix in | + | We divided up the PCR mix in 6 PCR tubes and added in each one a different clone from the transformed culture. |
| − | The picked colonies were | + | The picked colonies were spread on a Petri dish with LB + Ampicillin (50µg/mL) + X-Gal/IPTG (1/1000). |
Results : | Results : | ||
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!pPS16_002 | !pPS16_002 | ||
|- | |- | ||
| − | !''' | + | !'''Band Size (bp)''' |
|960 | |960 | ||
|} | |} | ||
| − | + | The electropheresis on agarose gel showed absolutely no PCR products. | |
| − | The electropheresis on agarose gel showed absolutely no PCR products. | + | pPS16_002 transformation from the 19/07/2016 was spread (50µL) on two Petri dishes LB + Ampicillin (50µg/mL) + X-Gal/IPTG (1/1000). |
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====High Fidelity Q5 PCR of transformed DH5α with pPS16_004 and pPS16_007 ==== | ====High Fidelity Q5 PCR of transformed DH5α with pPS16_004 and pPS16_007 ==== | ||
''By Laetitia'' | ''By Laetitia'' | ||
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The PCR was performed following the usual [[Team:Paris_Saclay/Experiments#Polymerase_chain_reaction|protocol]]. | The PCR was performed following the usual [[Team:Paris_Saclay/Experiments#Polymerase_chain_reaction|protocol]]. | ||
8 tubes were done: 5 clones of pPS16_004 and 3 clones of pPS16_007 | 8 tubes were done: 5 clones of pPS16_004 and 3 clones of pPS16_007 | ||
| − | The | + | The Tm was at 60°C |
| − | ====Glycerol stocks for DH5α transformed with | + | [[File:T--Paris_Saclay--160722_Visualisation_PCRpPS16_002_7.JPG|400px|thumb|right|Migration of pPS16_002 and pPS16_007]] |
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| + | ====Glycerol stocks for DH5α transformed with pcl_TAA, pcl_TAG and pcl_Tq ==== | ||
''By Laetitia'' | ''By Laetitia'' | ||
8 stocks were made: | 8 stocks were made: | ||
2 clones (Cl 1 and Cl 2) : | 2 clones (Cl 1 and Cl 2) : | ||
| − | * Cl1 and Cl2 of | + | * Cl1 and Cl2 of pcl_TAA |
| − | * Cl1 and Cl2 of | + | * Cl1 and Cl2 of pcl_TAG |
| − | * Cl1 and Cl2 of | + | * Cl1 and Cl2 of pcl_Tq |
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* 1mL of liquid culture | * 1mL of liquid culture | ||
* 500 μL of glycerol 60% | * 500 μL of glycerol 60% | ||
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{{Team:Paris_Saclay/notebook_footer}} | {{Team:Paris_Saclay/notebook_footer}} | ||
Latest revision as of 15:56, 9 October 2016
