Difference between revisions of "Team:Paris Saclay/Notebook/July/26"
(→High fidelity PCR on bacteria transformed with pPS16_001, pPS16_003, pPS16_005 and pPS16_009) |
(→High fidelity PCR on bacteria transformed with pPS16_001, pPS16_003, pPS16_005 and pPS16_009) |
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''By Alice'' | ''By Alice'' | ||
| − | PCR peformed on [[Team:Paris_Saclay/Notebook/July/25#PCR_1.1_3.1_GFP|July 25]] with bacteria containing plasmids pPS16_001, pPS16_005 and pPS16_009 did not give us expected results. We performed again this PCR, adding bacteria transformed with pPS16_003 | + | PCR peformed on [[Team:Paris_Saclay/Notebook/July/25#PCR_1.1_3.1_GFP|July 25]] with bacteria containing plasmids pPS16_001, pPS16_005 and pPS16_009 did not give us expected results. We performed again this PCR, adding bacteria transformed with pPS16_003 (clone 3) that got good sequencing results. The PCR with Q5® High-Fidelity 2X Master Mix was performed on these clones following [[Team:Paris_Saclay/Experiments#Q5PCR|this protocol]]. Specific [[Team:Paris_Saclay/Experiments#primers|primers]] were used for each plasmids (table below). Annealing temperature calculated was 72°C. After amplification, 1 µL of loading dye was added to 5µL of each PCR products. Then 5 µL of this mix were put into a well of the gel. A last well contained 10µL of DNA ladder. PCR products were migrated 25min at 100V. |
PCR products expected were : | PCR products expected were : | ||
| Line 23: | Line 23: | ||
|iPS120 | |iPS120 | ||
|960 | |960 | ||
| + | |- | ||
| + | |pPS16_003 | ||
| + | |2.1 | ||
| + | |iPS123 | ||
| + | |iPS124 | ||
| + | |1023 | ||
|- | |- | ||
|pPS16_005 | |pPS16_005 | ||
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|862 | |862 | ||
|} | |} | ||
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Revision as of 10:10, 26 July 2016
