The PCR-based KASP™ (i.e. Kompetitive Allele Specific PCR) genotyping assay is a homogeneous, fluorescence (FRET) based assay that enables accurate bi-allelic discrimination of known SNPs and InDels. Unlike other PCR based genotyping assays, KASP requires no labelling of the target-specific primers / probes. KASP allows for great flexibility for assay design, which translates into a higher overall success rate and the unique ability to genotype large / mega InDels.
A key feature of KASP technology is the use of a universal FRET cassette reporter system that eliminates the need for dual-labelled probes. The two allele-specific forward primers each have a tail sequence that corresponds with one of two FRET cassettes: one label with FAM dye and the other with HEX dye. This technology gives KASP the option to be ran at miniaturised reaction volumes (10 µL in 96-well PCR plates, 5 µL in 384-well PCR plates; 1 µL in 1536-well PCR plates) and delivers the benefit of individual PCR reactions.
KASP is very well suited for the range of 1 – 100 markers.