Site direct mutagenesis was
used to create the two active site mutants of the E.
coli exonuclease ε. The methionine 18 to alanine
(ε18) was created using the forward primer 5′-GAAACCACCGGTGCGAACCAGATTGGTGCG-3′
and reverse primer 5′-CGCACCAATCTGGTTCGCACCGGTGGTTTC-3.′
The valine 65 to alanine mutation (ε65) was created
using the forward 5′-GGAAGCCTTTGGCGCACATGGTATTGCCGATG-3′
and reverse primer 5′-CATCGGCAATACCATGTGCGCCAAAGGCTTCC-3.′
used to create the two active site mutants of the E.
coli exonuclease ε. The methionine 18 to alanine
(ε18) was created using the forward primer 5′-GAAACCACCGGTGCGAACCAGATTGGTGCG-3′
and reverse primer 5′-CGCACCAATCTGGTTCGCACCGGTGGTTTC-3.′
The valine 65 to alanine mutation (ε65) was created
using the forward 5′-GGAAGCCTTTGGCGCACATGGTATTGCCGATG-3′
and reverse primer 5′-CATCGGCAATACCATGTGCGCCAAAGGCTTCC-3.′
