Skip to main content

Table 1 Mutations, primers and restriction enzyme recognition site added or removed.

From: A new approach to 'megaprimer' polymerase chain reaction mutagenesis without an intermediate gel purification step

Template

Mutation

Mutagenic primer sequence

Megaprimer (bp)

Enzyme

E. coli KARI

E213D

5'-GTCCTTCGTTGCGGAcAAATCTGACCTG-3' (F)

968

Nil

A. thaliana AHAS Regulatory subunit

N346H

5'-CGAAGGGGgTACcaTATCCAGAGC-3' (F)

705

KpnI

S. cerevisiae AHAS Regulatory subunit

D224A

5'-GGTCAAGTTGGTGATAgCGTT-3' (R)

805

EcoRV

S. cerevisiae AHAS Catalytic subunit

M354V

5'-CAACCGTGCAcACCAAGCATATCC-3' (R)

1127

ApaL1

  1. In the primer sequences shown, bases that differ from the wild-type are shown in lowercase and the altered codon is underlined. Boxes enclose introduced restriction enzyme recognition sites, while strikethrough shows a restriction enzyme recognition site that is removed. The designations (F) and (R) indicate forward and reverse primers, respectively.