User:Christine Doan/Notebook/Identifying Kinase Substrates of PfCK2/2014/03/11: Difference between revisions
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* Ran cleanups on gel, look good | * Ran cleanups on gel, look good | ||
* Can re-PCR to amplify >70 and use those products for cloning into pET-30 | * Can re-PCR to amplify >70 and use those products for cloning into pET-30 | ||
** 50ul GoTaq PCR rection | |||
*** 25ul GoTaq Master Mix | |||
*** 1ul forward primer | |||
*** 1ul reverse primer | |||
*** 1-5ul DNA template (depending on what kind of DNA, concentration, end goal, etc) | |||
*** Nuclease-Free Water up to 50ul | |||
** PCR | |||
*** 95C 2mins denature | |||
*** 95C 30sec denature | |||
*** 39C 1min anneal | |||
*** 68C 5min extend | |||
*** 68C 10min extend | |||
*** 4C hold | |||
* T4 treatment for cloning requires 0.2 picamole | * T4 treatment for cloning requires 0.2 picamole | ||
Revision as of 12:50, 11 March 2014
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