Sensitivity and specificity of high-resolution melting analysis in screening unknown SNPs and genotyping a known mutation

文献类型： 外文期刊

第一作者： Ye, M. H.

作者： Ye, M. H.;Chen, J. L.;Zhao, G. P.;Zheng, M. Q.;Wen, J.;Ye, M. H.

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关键词： A-FABP/CAPNl I chicken;HRMA;SSCP analysis

期刊名称：ANIMAL SCIENCE PAPERS AND REPORTS （ 影响因子：1.078； 五年影响因子：1.097 ）

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收录情况： SCI

摘要： High-resolution melting analysis (HRMA) was used to screen potential SNPs in the exons of chicken CAPNl (mu-calpain/large subunit) gene. A total of 312 DNA samples from Beijing-you chickens were used for detection. Twelve pairs of primer were designedto amplify twelve different exons and SNPs were detected in five of them. HRMA was also compared with PCR-SSCP analysis for genotyping of a known SNP site in the chicken adipocyte fatty acid binding protein gene (A-FABP). Amplicons of 275-bp fragment, bracketing the polymorphic site, were grouped by PCR-SSCP into three genotypes designated as CC, TT and CT. Small amplicons (56 bp) within the 275-bp fragments were designed to maximize the 7m difference between homozygotes and to genotype all possible three genotypes after a single melting analysis successfully. Results from different methods were cross-validated and sequencing results from randomly selected heterozygotes and homozygotes confirmed the specificity of HRM technique. The full consistency proved that HRMA was a useful tool for rapid, close-tube genotyping of polymorphic sites. It has great potential for SNPs detection and scanning especially on a large scale.

分类号： S8

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