IGEM:Groningen/2009/Notebook/iGEM 2011/2011/07/08: Difference between revisions
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== | ==*8th of July== | ||
Colony PCR of transformants to check if any transformant contains the plasmid with cI-LVA and LasR-LVA. | |||
20μl total volume was used and the PCR was done with taq polymerase. | |||
PCR program: | |||
Denaturation: 95 °C for ten minutes | |||
Cycle (33×): | |||
Denaturation: 95°C for 30 seconds | |||
Annealing: 60°C for 30 seconds | |||
Extension: 72°C for 2 minutes | |||
Final extension: 72°C for ten minutes | |||
Store at 4 °C infinite | |||
PCR with the new pBAD primers (dissolved and dilution made by me) | |||
PCR was done in total colume of 50μl and with tag and Pfu polymerase (ratio 6:1) so Pfu can correct the mistakes that taq made | |||
Denaturation: 94°C for ten minutes | |||
Cycle (30×) | |||
Denaturation: 94°C for 30 seconds | |||
Annealing: 62°C and 65°C for 30 seconds | |||
Extension: 72°C for 2 minutes | |||
Final extension: 72°C for ten minutes | |||
Store at 4 °C infinite | |||
Check them on agarose gel 1% (TBE). | |||
After that: Clean up the pBAD samples and nanodrop the samples | |||
Nanodrop results: | |||
Revision as of 03:16, 8 July 2011
 iGEM Project name 1
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*8th of JulyColony PCR of transformants to check if any transformant contains the plasmid with cI-LVA and LasR-LVA. 20μl total volume was used and the PCR was done with taq polymerase. PCR program: Denaturation: 95 °C for ten minutes Cycle (33×): Denaturation: 95°C for 30 seconds Annealing: 60°C for 30 seconds Extension: 72°C for 2 minutes Final extension: 72°C for ten minutes Store at 4 °C infinite PCR with the new pBAD primers (dissolved and dilution made by me) PCR was done in total colume of 50μl and with tag and Pfu polymerase (ratio 6:1) so Pfu can correct the mistakes that taq made Denaturation: 94°C for ten minutes Cycle (30×) Denaturation: 94°C for 30 seconds Annealing: 62°C and 65°C for 30 seconds Extension: 72°C for 2 minutes Final extension: 72°C for ten minutes Store at 4 °C infinite Check them on agarose gel 1% (TBE). After that: Clean up the pBAD samples and nanodrop the samples Nanodrop results:
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