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Team:HokkaidoU Japan/Notebook/plastic Week 7
From 2012.igem.org
Contents |
August 16th
PCR
A PCR of pGEM to get linear PhaA and PhaB using Laury primer and X-PhaA-up-F, S-PhaA-dw-R, X-PhaB-up-F, S-PhaB-dw-R.
| pGEM(pGEM 1ul + DW 9ul) | 1 ul |
| up primer(10mM) | 1 ul |
| down primer(10mM) | 1 ul |
| 25mM MgSO4 | 3 ul |
| 2mM sNTPs | 5 ul |
| KOD plus neo | 1ul |
| 10xPCR buffer | 5 ul |
| DW | 33 ul |
PCR:60sec, 30 cycle 1h8m
Did not work well?
August 17th
Electrophoresis
We confirmed success of PCR of yesterday by electrophoresis.
Gel extraction
The plasmids done PCR were extracted from TBE gel, and then we got 50 ul DNA solutions.
August 18th
Calculation of concentration for digestion
