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Team:HokkaidoU Japan/Notebook/plastic Week 13
From 2012.igem.org
Contents |
September 24th
Single colony isolation
We got few colonies from the plate. The colonies were isolated to another plate.
September 25th
Liquid culture
We got colonies on the plate, and started liquid culture.
PCR
Did PCR for promoter of R.eutropha from pGEM to ligate with RBS-PhaC-RBS-PhaA-RBS-PhaB-dT on pSB1A2.
Digestion
Digested the promoter done PCR and RBS-PhaA on pSB1A2 with XbaI and SpeI, and plasmid of RBS-PhaC-RBS-PhaA-RBS-PhaB-dT on pSB1A2 with XbaI.
Gel extraction
We confirmed succession of digestion by electrophoresis. And then DNA were extracted from TBE gel.
Ethanol precipitation
The digested DNAs were condensed by Ethanol precipitation.
Ligation
Ligated the promoter with RBS-PhaC-RBS-PhaA-RBS-PhaB-dT and with pSB1A2.
Transformation
Transformed each ligation product into JM109, and incubated.
September 26th
Colony PCR
Colony PCR for colonies transformed at 25th.
