A novel technique for detecting single nucleotide polymorphisms by analyzing consumed allele-specific primers.
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 ABO 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.
Department of Forensic Medicine, Yamagata University School of Medicine, Japan.
G
G
Watanabe
K
K
Umetsu
I
I
Yuasa
M
M
Sato
M
M
Sakabe
E
E
Naito
H
H
Yamanouchi
T
T
Suzuki
D000017:ABO Blood-Group System / Q000145:classification / Q000235:genetics*; D000483:Alleles; D004247:DNA / Q000032:analysis*; D017931:DNA Primers; D005838:Genotype; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D020641:Polymorphism, Single Nucleotide
D000017:ABO Blood-Group System; D017931:DNA Primers; D004247:DNA