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Table 2 PCR reaction components.

From: Simple and efficient site-directed mutagenesis using two single-primer reactions in parallel to generate mutants for protein structure-function studies

 

Reaction 1

Reaction 2

Template plasmid DNA

~500 ng

~500 ng

Forward primer

40 pmol

-

Reverse primer

-

40 pmol

MgCl2

0.2 mM

0.2 mM

dNTPs

0.2 mM

0.2 mM

Pwo Master DNA polymerase

1.25 U

1.25 U

Tris buffer, pH 7.5

10 mM

10 mM

Final volume

25 μl

25 μl