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Cost‐effective interrogation of single nucleotide polymorphisms using the mismatch amplification mutation assay and capillary electrophoresis
Author(s) -
Price Erin P.,
Matthews Molly A.,
Beaudry Jodi A.,
Allred Jonathan L.,
Schupp James M.,
Birdsell Dawn N.,
Pearson Talima,
Keim Paul
Publication year - 2010
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201000379
Subject(s) - interrogation , capillary electrophoresis , chromatography , microbiology and biotechnology , single nucleotide polymorphism , capillary action , chemistry , genetics , biology , materials science , genotype , gene , history , archaeology , composite material
The ability to characterize SNPs is an important aspect of many clinical diagnostic, genetic and evolutionary studies. Here, we designed a multiplexed SNP genotyping method to survey a large number of phylogenetically informative SNPs within the genome of the bacterium Bacillus anthracis. This novel method, CE universal tail mismatch amplification mutation assay (CUMA), allows for PCR multiplexing and automatic scoring of SNP genotypes, thus providing a rapid, economical and higher throughput alternative to more expensive SNP genotyping techniques. CUMA delivered accurate B. anthracis SNP genotyping results and, when multiplexed, saved reagent costs by more than 80% compared with TaqMan real‐time PCR. When real‐time PCR technology and instrumentation is unavailable or the reagents are cost‐prohibitive, CUMA is a powerful alternative for SNP genotyping.