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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