Difference between revisions of "Team:Paris Saclay/Notebook/August/9"
(→Phusion PCR on pPS16_006 and pPS16_007) |
(→pUC19 digestion with HincII) |
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The PCR was carried out following the usual [[Team:Paris_Saclay/Experiments#phusion|protocol]] adapted to 50µL and with a TM at 72°C. The specific [[Team:Paris_Saclay/Experiments#primers|primers]] for each parts were using. The products were put to migrated on a 0.8%agarose gel with BET. | The PCR was carried out following the usual [[Team:Paris_Saclay/Experiments#phusion|protocol]] adapted to 50µL and with a TM at 72°C. The specific [[Team:Paris_Saclay/Experiments#primers|primers]] for each parts were using. The products were put to migrated on a 0.8%agarose gel with BET. | ||
| − | ====pUC19 digestion with HincII ==== | + | ====pUC19 digestion with HincII enzyme==== |
''By Alice'' | ''By Alice'' | ||
| − | 5 µL of pUC19 plasmids were digested with 5µL of tango buffer 10X, 38µL of sterile water, and 1 µL of HincII enzyme. The mix was incubated at 37°C for 1 hour. After incubation, 1µL of HincII enzyme was added again, and the mix was incubated 1 hour again. | + | |
| + | 5 µL of pUC19 plasmids were digested with 5µL of tango buffer 10X, 38µL of sterile water, and 1 µL of HincII enzyme. The mix was incubated at 37°C for 1 hour. After incubation, 1µL of HincII enzyme was added again, and the mix was incubated 1 hour again. Digestion products were migrated on a gel on [[Team:Paris_Saclay/Notebook/August/10#August_10|August 10]]. | ||
===pUC19 ligation with gBlocks ATG linker RFB, detection, St sgRNA, ATG linker FKBP, Nm sgRNA, 1.2 and 4.2=== | ===pUC19 ligation with gBlocks ATG linker RFB, detection, St sgRNA, ATG linker FKBP, Nm sgRNA, 1.2 and 4.2=== | ||
Revision as of 13:09, 10 August 2016
