The PCR-based KASP™ (Kompetitive Allele Specific PCR) genotyping assay is a homogeneous, fluorescence (FRET) based technique that enables precise bi-allelic discrimination of known SNPs and InDels. Unlike other PCR-based genotyping assays, KASP does not require labeling of the target-specific primers/probes.
This characteristic grants KASP exceptional flexibility in assay design, resulting in a higher overall success rate and the unique capability to genotype large/mega InDels.
KASP technology
A pivotal aspect of KASP technology is its utilization of a universal FRET cassette reporter system, negating the necessity for dual-labeled probes. The two allele-specific forward primers each possess a tail sequence corresponding to one of two FRET cassettes: one labeled with FAM dye and the other with HEX dye.
This technological advancement enables KASP to be performed at miniaturized reaction volumes (10 µL in 96-well PCR plates, 5 µL in 384-well PCR plates; 1 µL in 1536-well PCR plates) while delivering the advantage of individual PCR reactions.
KASP is highly adaptable for a spectrum of 1–100 markers and serves as an excellent solution for screening plant variants.
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