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Polymerase Chain Reaction:
10 X buffer:
100 mM Tris/HCl pH 8.3
0.5 M KCl
15 mM MgCl2
10 x dNTP stock solution: 2.5 mM each. (dATP, dCTP, dGTP,dTTP)
Taq Polymerase: 2.5 units/ 100 ul reaction
Primers 50 picomoles
Template >= 10 ng/ul
Final Volume = 50 ul.
Determine Tm of primers. Set initital annealing temperature at Tm – 5O
C using the primer with the lowest Tm as the guide.
Promega PCR guide: http://www.promega.com/amplification/prodguide/promega.html#Contents
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