Rapid and Efficient Site-directed Mutagenesis by the Single-tube Megaprimer PCR Method

2006 ◽  
Vol 2006 (1) ◽  
pp. pdb.prot3467 ◽  
Author(s):  
Joseph Sambrook ◽  
David W. Russell
Keyword(s):  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Single Tube ◽  
Pcr Method

scholarly journals Use of site-directed mutagenesis of allele-specific PCR primers to identify the GSTM1 A, GSTM1 B, GSTM1 A,B and GSTM1 null polymorphisms at the glutathione S-transferase, GSTM1 locus

1993 ◽  
Vol 295 (1) ◽  
pp. 313-315 ◽  
Author(s):  
A A Fryer ◽  
L Zhao ◽  
J Alldersea ◽  
W R Pearson ◽  
R C Strange
Keyword(s):  
Pcr Primers ◽  
Single Step ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Glutathione S Transferase ◽  
Specific Pcr ◽  
Target Dna ◽  
Allele Specific ◽  
Pcr Method ◽  
Allele Specific Pcr

We describe the identification of the GSTM1 null, GSTM1 A, GSTM1 B and GSTM1 A,B polymorphisms at the glutathione S-transferase GSTM1 locus using a single-step PCR method. Target DNA was amplified using primers to intron 6 and exon 7 with site-directed mutagenesis being used to introduce a restriction site in DNA amplified from GSTM1 *A, thereby allowing differentiation of this allele and GSTM1 *B. The accuracy of this approach in identifying the GSTM1 A, GSTM1 B, GSTM1 A,B and GSTM1 null polymorphisms was confirmed by comparison with, firstly, an established PCR method that distinguishes GSTM1 *0 homozygotes from individuals with the other GSTM1 genotypes and, secondly, GSTM1 phenotypes determined using chromatofocusing.

scholarly journals Efficient site-directed mutagenesis using an overlap extension-PCR method for expressing Mycoplasma hyopneumoniae genes in Escherichia coli

2009 ◽  
Vol 79 (1) ◽  
pp. 101-105 ◽  
Author(s):  
Simone Simionatto ◽  
Silvana B. Marchioro ◽  
Vanessa Galli ◽  
Tessália D. Luerce ◽  
Daiane D. Hartwig ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Mycoplasma Hyopneumoniae ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Overlap Extension Pcr ◽  
Overlap Extension ◽  
Pcr Method
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