Team:HokkaidoU Japan/Notebook/plastic Week 7

From 2012.igem.org

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==August  18th==
==August  18th==

Revision as of 11:54, 14 September 2012

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

Gel extraction


August 18th

Calculation of concentration for digestion