AcycloPrime: a novel method for SNP analysis using fluorescence polarization

Single nucleotide polymorphisms (SNPs) are the most common type of genetic variation between individuals of a species and are therefore thought to be responsible for a large part of individual phenotypic variation. It has been estimated that a SNP may occur every 100-300 bases in the human genome. Research on human SNPs is expected to facilitate genetic mapping studies that may lead to a better understanding of the genetic basis for complex diseases and individual variation in drug metabolism. We have developed a novel assay for the identification of known SNPs using primer extension with the novel AcycloTerminators and a new thermostable polymerase, AcycloPol, in a homogeneous fluorescence polarization (FP) format. All assay steps can be performed in the same well of either a 384- or 96-well PCR-compatible microplate. FP provides several advantages, including simplicity and low reagent cost. The homogeneous assay format eliminates any need for separation or washing steps and is amenable to automation.