A Novel Multiplex PCR-Based Solution for SNP Genotyping

See Jana Kent, PhD present a novel, one-step SNP genotyping methodology with high discrimination power and scalable multiplex capability. This simple, PCR-based system relies on allele-specific iteration of previously reported Cooperative Primers. The CoPrimer genotyping assay involves two allele-specific and one locus-specific oligonucleotides per SNP. Each allele-specific CoPrimer possesses a Priming sequence hybridizing at the SNP locus, and a Capture probe labeled with a fluorophore that is spectrally distinct for each allele. Multiplexing capabilities of CoPrimers allow for interrogation of several SNPs in one tube - thus saving reagents, time and significantly reducing the amount of the DNA sample needed. The CoPrimer genotyping system has been tested with PCR Master Mixes from several manufacturers. Best performance was observed with BHQ® Probe Master Mix (LGC), which was also the only Master Mix that allowed for SNP multiplexing without loss of PCR efficiency.

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