Denise Yu: Difference between revisions

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5. 72C for 5 minutes
5. 72C for 5 minutes
[[Image:PCR-gel-11-11-2008.jpg]]

Revision as of 23:54, 11 November 2008

Hot Start PCR (11/10/08)


Reagent

-10x Thermo-Start Standard Buffer (5 microliters)

-20mM dNTP Mix (2 microliters)

-25mM MgCl2 (3 microliters)

-Primer Mix (2.5 microliters)

-Template DNA 10 ng

-Thermo-Start DNA Polymerase (.25 microliters)

-double-distlled, sterile H2O (up to 50 microliters)


Thermal Cycle

Enzyme Activation

1. 95C for 15 minutes

Denaturation

2. 92-95C for 10-30 seconds

Annealing

3.50-65C for 30 seconds, repeart 25-30

Extension:

4.72C for 30 seconds

Final Extension:

5. 72C for 5 minutes

File:PCR-gel-11-11-2008.jpg