The KASP genotyping assay utilises a unique form of competitive allele-specific PCR that delivers extremely high levels of assay robustness and accuracy. Originally developed and patented by KBioscience the KASP genotyping assay is a singleplex PCR-based assay that enables highly accurate bi-allelic scoring of SNPs (single nucleotide polymorphisms) & InDels (Insertions and Deletions) at specific loci across a wide range of genomic DNA samples, including those of complex genomes.
Available in reagent kit form for in-lab-use, or as a genotyping service through our genomic service laboratories in Europe and North America, the KASP genotyping assay delivers:
Assembled as a homogeneous assay in 96, 384, or 1536-well PCR plates, bi-allelic discrimination is achieved through the competitive binding of two allele-specific forward primers, each with unique tail sequences that correspond with two universal FRET (fluorescence resonant energy transfer) cassettes; one labeled with FAM™ dye and the other with HEX™ dye.
Importantly the allele-specific primers for each assay require no labelling, which gives the KASP genotyping assay a distinct cost advantage relative to other genotyping methodologies. And with far greater flexibility in terms of primer selection and assay design, the KASP genotyping assay also outperforms most in terms of the range of polymorphisms that can be scored, including SNP’s in high variability regions and Mega InDels. Through our service labs we often delight customers by getting the KASP genotyping assay to work when other methodologies they’ve tried have failed.
There are two reagent components to the KASP genotyping assay:
Running a KASP genotyping assay is easy. Simply add the assay specific KASP Primer mix and the universal KASP Master mix to your DNA sample in either a 96, 384, or 1536-well PCR plate, seal the plate, run your thermal cycling reaction, and then conduct an end-point read using either a RT-PCR machine, or separate fluorescent plate reader. No separation steps are required.
KASP chemistry can be used with any standard thermal cycler. Equivalent results have been demonstrated on traditional peltier block-based thermal cyclers and our own Hydrocycler line of water bath-based thermal cyclers.