KASP genotyping technology
The patented PCR-based KASP™ 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, giving it a clear cost advantage. It also allows greater 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 LGC's PCR-based KASP technology is the use of a universal FRET cassette reporter system that eliminates the need for costly dual-labelled probes. LGC's allele-specific forward primers each have a proprietary 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 a clear cost advantage, and when combined with miniaturized reaction volumes (10 µL in 96-well PCR plates, 5 µL in 384-well PCR plates; 1 µL in 1536-well PCR plates), delivers an exceptionally accurate, flexible, and cost-effective genotyping solution. Bi-allelic discrimination is achieved through the competitive binding of the two allele-specific forward primers.
Why use KASP?
The simplicity of the KASP assay design yields clear end-user advantages including:
- Superb accuracy and performance
- Accuracy: >99.8% based on independent assessment
- Industry leading SNP & InDel assay conversion rate (>90%)
- Tremendous flexibility
- Flexible primer design which increases the rate of successful assay development
- Supports low-, medium- and high-throughput studies and individual repeat assays
- Compatibility with a range of liquid handling systems and thermal cyclers, signal can be read on most FRET-capable plate readers and qPCR instruments.
- Breakthrough cost savings
- KASP uses a universal reporting system where the labelled components are present in the KASP Master mix
- Eliminates the need for expensive labelled assay-specific primers or probes
- Requires only 10 ng DNA per sample per SNP (based on human genome size)
- Low reaction volumes keep reagent costs to a minimum
- Cost benefits enable you to perform more assays overall, improving the quality of your data.
The KASP genotyping technology is based on an open format PCR workflow. There is no need to buy new expensive equipment to be able to run KASP in your own laboratory; completed KASP reactions can be read on most FRET-capable plate readers and qPCR instruments.