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A novel technique for detecting single nucleotide polymorphisms by analyzing consumed allele‐specific primers
Author(s) -
Watanabe Gotaro,
Umetsu Kazuo,
Yuasa Isao,
Sato Michihiko,
Sakabe Munechika,
Naito Emiko,
Yamanouchi Haruo,
Suzuki Tsuneo
Publication year - 2001
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/1522-2683(200102)22:3<418::aid-elps418>3.0.co;2-8
Subject(s) - genotyping , primer (cosmetics) , single nucleotide polymorphism , polymerase chain reaction , biology , genetics , variants of pcr , snp genotyping , nucleotide , typing , allele , in silico pcr , molecular inversion probe , primer dimer , microbiology and biotechnology , genotype , multiplex polymerase chain reaction , gene , chemistry , organic chemistry
We present a simple and rapid polymerase chain reaction (PCR)‐based technique, termed consumed allele‐specific primer analysis (CASPA), as a new strategy for single nucleotide polymorphism (SNP) analysis. The method involves the use of labeled allele‐specific primers, differing in length, with several noncomplementary nucleotides added in the 5'‐terminal region. After PCR amplification, the amounts of the remaining primers not incorporated into the PCR products are determined. Thus, nucleotide substitutions are identified by measuring the consumption of primers. In this study, the CASPA method was successfully applied to AB0 genotyping. In the present method, the allele‐specific primer only anneals with the target polymorphic site on the DNA, so it is not necessary to analyze the PCR products. Therefore, this method is only little affected by modification of the PCR products. The CASPA method is expected to be a useful tool for typing of SNPs.