Open AccessApplication of Tm-shift genotyping method in genetic studies
Author(s) -
Fang Yuan,
Jin Xu,
Li Ji,
Li-Juan Fei,
Panpan Li,
Lina Zhang
Publication year - 2012
Publication title -
yichuan
Language(s) - English
Resource type - Journals
ISSN - 0253-9772
DOI - 10.3724/sp.j.1005.2012.01484
Subject(s) - genotyping , single nucleotide polymorphism , melting curve analysis , genotype , snp genotyping , genetics , biology , repeatability , polymerase chain reaction , chromatography , chemistry , gene
Melting Temperature shift (Tm-shift) is a new genotyping method. With two GC-rich tails of unequal length combined to 5'-terminal of allele-specific primers, genotypes can be determined by the distinct Tms of the PCR products with inspection of a melting curve on the real-time PCR machine. In this study, 29 SNPs were genotyped with 2 048 samples by using Tm-shift genotyping method, and the results were assessed by success rate, consistent rate, and accuracy. The results indicated that among 29 SNPs, 27 SNPs could be genotyped by Tm-shift. In other words, the success rate was 93.1%. The accuracy confirmed by direct sequencing was 100%. The consistency was 100% with 3 control samples, and 97% from a replication study in 100 samples. Thus, Tm-shift is a genotyping method with advantages including low cost, high accuracy, stability, reliability, flexible throughput, and easy operation, which can be applied to genetic studies widely.