Tm-shift基因分型方法在遗传学中的应用

doi:10.3724/SP.J.1005.2012.01484

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

Tm-shift基因分型方法在遗传学中的应用

袁芳¹, 徐进^1,2, 季林丹^1,2, 费丽娟¹, 刘盼盼¹, 张莉娜¹

1. 宁波大学医学院, 宁波 315211 2. 中国科学院昆明动物研究所-香港中文大学生物资源与疾病分子机理联合实验室, 昆明 650223

收稿日期:2012-08-27 修回日期:2012-10-10 出版日期:2012-11-20 发布日期:2012-11-25
通讯作者: 张莉娜 E-mail:zhanglina@nbu.edu.cn
基金资助:
浙江省自然科学基金项目(编号：(Y2100857))、浙江省教育厅重点科研项目(编号：Z201017918)和宁波市自然科学基金项目(编号：2012A610237, 2011A610037)资助

Application of Tm-shift genotyping method in genetic studies

YUAN Fang¹, XU Jin^1,2, JI Lin-Dan^1,2, FEI Li-Juan¹, LIU Pan-Pan¹, ZHANG Li-Na¹

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

摘要/Abstract

摘要： Melting Temperature shift(Tm-shift)是一种新的基因分型方法, 主要通过在两条特异性引物5′端加入不同长度的GC序列, PCR扩增后根据熔解曲线中产物Tm值的差异来完成分型。文章建立了Tm-shift法对2 048份样品的29个SNP进行分型, 通过分型成功率、重复检测一致率、测序验证准确度综合评价分型效果。结果显示, 29个SNP中有27个可以采用本方法分型, 分型成功率为93.1%。测序验证准确性达到100%。3种基因型阳性标准对照重复检测一致率为100%; 100个随机样品重复检测, 重复性为97%。因此, Tm-shift基因分型法是一种成本低廉、准确灵敏、稳定可靠、通量灵活、操作简便的基因分型方法, 可在遗传学研究中推广应用。

关键词: Tm-shift, 基因分型, 遗传学

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

袁芳，徐进，季林丹，费丽娟，刘盼盼，张莉娜. Tm-shift基因分型方法在遗传学中的应用[J]. 遗传, 2012, 34(11): 1484-1490.

YUAN Fang XU Jin JI Lin-Dan FEI Li-Juan LIU Pan-Pan ZHANG Li-Na. Application of Tm-shift genotyping method in genetic studies[J]. HEREDITAS, 2012, 34(11): 1484-1490.

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Tm-shift基因分型方法在遗传学中的应用

Application of Tm-shift genotyping method in genetic studies

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