Abstract

In order to promote the widespread application of single-nucleotide polymorphism (SNP)-based genotyping, a new method was developed and called target SNP-seq which combined the advantages of multiplex polymerase chain reaction (PCR) amplification and high throughput sequencing on Illumina X Ten platform. Compared with kompetitive allele-specific PCR (KASP), microchips, and genotyping by sequencing (GBS), target SNP-seq uses perfect SNPs based on the analysis of variome (whole-genome sequence data of different accessions) and is flexible, cost-effective, and highly accurate for genotyping middle-scale SNPs. It could genotype hundreds of SNPs in massive DNA samples within 3days at the cost of $7 for each DNA sample. The high efficiency and low cost of target SNP-seq make it more competitive than current SNP genotyping methods, and it has excellent potential for application in genetic research, as well as in promoting plant-breeding processes in the near future.

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