Difference between revisions of "Team:Paris Saclay/Notebook/September/16"
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| + | {{Team:Paris_Saclay/notebook_header}} | ||
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= Friday 16<sup>th</sup> September= | = Friday 16<sup>th</sup> September= | ||
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===Visualization=== | ===Visualization=== | ||
| − | ====Colony PCR of 16 clones containing | + | ====Colony PCR of 16 clones containing FKBP - GFP 10 in pSB1C3 (pPS16_019)==== |
''By Mahnaz'' | ''By Mahnaz'' | ||
Colonies were obtained for the Gibson done the 15th September. A colony PCR was done for 16 clones from the 15th September. For that purpose, 16 were screened and used for the usual [[Team:Paris_Saclay/Experiments##Polymerase_chain_reaction|protocol]] of Colony PCR. The same colonies were also plated on Petri dish containing solid LB + 30 µg/mL Cm and liquid cultures (LB + 30 µg/mL Cm) were made from these clones. Both petri dishes and liquid cultures were grown at 37°C. | Colonies were obtained for the Gibson done the 15th September. A colony PCR was done for 16 clones from the 15th September. For that purpose, 16 were screened and used for the usual [[Team:Paris_Saclay/Experiments##Polymerase_chain_reaction|protocol]] of Colony PCR. The same colonies were also plated on Petri dish containing solid LB + 30 µg/mL Cm and liquid cultures (LB + 30 µg/mL Cm) were made from these clones. Both petri dishes and liquid cultures were grown at 37°C. | ||
| − | For each clones contained in 20 μl water, | + | For each clones contained in 20 μl water, 5.13 μL of the following mix were added : |
* 2.5 µL DreamTaq Buffer | * 2.5 µL DreamTaq Buffer | ||
* 0.5 µL of dNTPs (10mM) | * 0.5 µL of dNTPs (10mM) | ||
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|72°C | |72°C | ||
| − | | | + | |30 sec |
|- | |- | ||
|Final Extension | |Final Extension | ||
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|} | |} | ||
| − | + | [[Team:Paris_Saclay/Experiments#primers|Primers]] used were: | |
{| class="wikitable" | {| class="wikitable" | ||
|- | |- | ||
!Matrix | !Matrix | ||
| − | !Clones containing FKBP - GFP 10 in pSB1C3 | + | !Clones containing FKBP - GFP 10 in pSB1C3 (pPS16_19) |
|- | |- | ||
|Primers | |Primers | ||
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!Expected band size (bp) | !Expected band size (bp) | ||
|- | |- | ||
| − | |FKBP - GFP 10 in pSB1C3 | + | |FKBP - GFP 10 in pSB1C3 (pPS16_019) |
|757 | |757 | ||
|} | |} | ||
| − | + | [[File:T--Paris Saclay--Gel1118.png|400px|thumb|center|Result of the migration]] | |
| + | |||
| + | Some PCR products were at the good size. Clones 7, 8, 9 and 10 were selected and were grown at 37°C overnight. | ||
| + | |||
| + | {{Team:Paris_Saclay/notebook_footer}} | ||
Latest revision as of 17:06, 9 October 2016
