title: [Application of Tm-shift genotyping method in genetic studies].
abstract: 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.
authors.affiliation: Department of Preventive Medicine, Ningbo University, Ningbo, China. yuanfang40@163.com
authors.forename: Fang
authors.initials: F
authors.lastname: Yuan
authors.forename: Jin
authors.initials: J
authors.lastname: Xu
authors.forename: Lin-Dan
authors.initials: LD
authors.lastname: Ji
authors.forename: Li-Juan
authors.initials: LJ
authors.lastname: Fei
authors.forename: Pan-Pan
authors.initials: PP
authors.lastname: Liu
authors.forename: Li-Na
authors.initials: LN
authors.lastname: Zhang
mesh_terms: D060005:Genotyping Techniques; D008274:Magnesium / Q000494:pharmacology; D016133:Polymerase Chain Reaction; D020641:Polymorphism, Single Nucleotide; D044366:Transition Temperature
chemical_list: D008274:Magnesium