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Team:HokkaidoU Japan/Notebook/plastic Week 7
From 2012.igem.org
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====PCR==== | ====PCR==== | ||
Revision as of 01:23, 27 September 2012
Contents |
August 16th
PCR
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 1 ul + DW 9 ul) | 1 ul |
| up primer(10 mM) | 1 ul |
| down primer(10 mM) | 1 ul |
| 25 mM MgSO4 | 3 ul |
| 2 mM sNTPs | 5 ul |
| KOD plus neo | 1 ul |
| 10xPCR buffer | 5 ul |
| DW | 33 ul |
| Number | Degree | Second |
| 1 | 94 | 120 |
| 2 | 98 | 10 |
| 3 | 68 | 60 |
| 4 | 4 | HOLD |
Cycle:2~3 x 30
Did not work well?
August 17th
Electrophoresis
We confirmed success of PCR of August 16th by electrophoresis.
Gel extraction
The plasmids done PCR were extracted from TBE gel, and then we got 50 ul DNA solutions.
