Abstract

Single nucleotide polymorphism (SNP) can be detected by mining sequence databases or by using different singleplex or multiplex SNP genotyping platforms. Development of high-throughput genotyping molecular markers can be instrumental towards maximizing genetic gain. In this review we provide an overview of Kompetitive Allele Specific PCR (KASP) genotyping platform requirements and its application that might be helpful in KASP marker development. This literature further illustrates the possibilities to design KASP primers. Several research institutes routinely using KASP platform, producing in excess of humungous data points yearly for breeding cultivars and as well as for medical and commercial purposes. KASP genotyping technology offers cost effectiveness and high throughput molecular marker development platform. Conventional molecular markers can be converted into more robust and high throughput KASP markers. More than 2000 published references clearly show the popularity of KASP technology among the researchers.

Highlights

  • Development of molecular markers that predict the trait of interest with highest level of accuracy offers efficient tracking of superior genetic variations through breeding process bypassing phenotypic evaluation requirements

  • Most of the molecular markers developed for marker assisted selection (MAS) are diagnostic markers which forecast phenotype with varying degree of accuracy

  • Single nucleotide polymorphism (SNP) are abundant in genomes and they are amenable for high throughput detection

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Summary

Introduction

Development of molecular markers that predict the trait of interest with highest level of accuracy offers efficient tracking of superior genetic variations through breeding process bypassing phenotypic evaluation requirements. Primers create mutations into the target sequences during amplification and conjunction with the discovered SNP it results in formation of a unique restriction site Both CAPS and dCAPS are gel-based genotyping methods. Scoring of genotype is laborious, time consuming and might cause error while scoring for large set of population (Figure 1) To overcome these limitations to a considerable level, gel free high throughput single plex SNP genotyping platforms such as Kompetitive Allele Specific PCR (KASPTM) is more suitable (Neelam et al, 2013). Successful conversion of sequence-tagged site (STS) gel based marker for Lr21 into KASP assay has been reported with high throughput, accurate, inexpensive and less laborious (Neelam et al, 2013). KASP genotyping system used by Kusza and co-workers on 48 polymorphisms at different caprine loci and French Alpine and Saanen goat breeds found 94.81% assay success rate and found a high locus specific and accurate genotyping solution (Kusza et al, 2018)

KASP Requirements
Sampling Assayed
Thermal Cycling Conditions
10. Graphical Viewing of Genotyping Data
11. KASP Application
12. KASP vs Amplifluor and TaqMan
Findings
13. KASP Drawbacks and Alternative SNP Genotying Techniques
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