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
Forward primer
40 pmol
-
Reverse primer
MgCl2
0.2 mM
dNTPs
Pwo Master DNA polymerase
1.25 U
Tris buffer, pH 7.5
10 mM
Final volume
25 μl