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HEREDITAS ›› 2012, Vol. 34 ›› Issue (11): 1484-1490.doi: 10.3724/SP.J.1005.2012.01484

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Application of Tm-shift genotyping method in genetic studies

YUAN Fang1, XU Jin1,2, JI Lin-Dan1,2, FEI Li-Juan1, LIU Pan-Pan1, ZHANG Li-Na1   

  1. 1. Department of Preventive Medicine, School of Medicine, Ningbo University, Ningbo 315211, China 2. KIZ-CUHK Joint Laboratory of Bioresources and Molecular Research of Common Diseases, Kunming 650223, China
  • Received:2012-08-27 Revised:2012-10-10 Online:2012-11-20 Published:2012-11-25

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.

Key words: Tm-shift, genotyping, genetics