Team:HokkaidoU Japan/Notebook/plastic Week 7

From 2012.igem.org

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.