Homogeneous Allele-Specific PCR in SNP Genotyping

Allele-specific (AS) PCR amplification (1 –3 ) has been used in combination with gel based detection to genotype-specific polymorphisms. Until recently a major drawback of this method was that it was labor-intensive and without high-throughput instrumentation (4 ). The single nucleotide polymorphism (SNP) genotyping assay presented here combines AS PCR amplification with kinetic, realtime monitoring (5 –6 ). It is robust, rapid, inexpensive, and allows accurate measurement of allele frequencies in pools of DNA, facilitating large-scale gene mapping.