Denise Yu: Difference between revisions

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Reagent
Reagent
-10x Thermo-Start Standard Buffer (5 microliters)
-10x Thermo-Start Standard Buffer (5 microliters)
-20mM dNTP Mix (2 microliters)
-20mM dNTP Mix (2 microliters)
-25mM MgCl2 (3 microliters)
-25mM MgCl2 (3 microliters)

Revision as of 23:46, 10 November 2008

Hot Start PCR

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