One tube nested-PCR 在SNP 基因型分型中的应用 - 湖南农业大学 .PDF

() 2015 4 41 2 Journal of Hunan Agricultural University (Natural Sciences), Apr. 2015, 41(2):136–142 DOI:10.13331/ki.jhau.2015.02.005 One tube nested-PCR 在 SNP 基因型分型中的应用 1 1 2 1 2 (1. 723000 2. 723000)   摘 要nested–PCR PCR (SNP) nested–PCR SNP 1 Fgr 3 Pi–taPi9Pigm nested–PCR 4 PCR SNP DNA SNP 关 键 词nested–PCR 中图分类号Q789 文献标志码：A 文章编号：1007− 1032(2015)02−0136−07 Application of one tube nested-PCR on genotyping of SNP 1 1 2 1 2 Zhang Yu , Zhang Xiaojuan ,Wang Shengbao , Song Xiaoli , Li Xiaogang (1.School of Biological Sciences and Engineering, Shaanxi University of Technology, Hanzhong, Shaanxi 723000, China; 2.Hanzhong Institute of Agricultural Sciences, Hanzhong, Shaanxi 723000China)   Abstract: Based on common PCR, nested–PCR technology was developed to specially detect single nucleotide polymorphism (SNP). In this study, one tube nested–PCR technique was adopted to explore SNPs in four structural genes of rice, there were Fgr, a rice aroma gene and Pi–ta, Pi9 and Pigm, three rice blast resistance genes. Primers with effectively marked at targeting SNP sites were designed and four primers were mixed in one tube to amplify DNA fragments harboring mutation sites. In addition, reaction conditions including primer concentration and PCR program were optimized to identify different SNP genotypes in one step. Therefore, the one tube nested–PCR provided a feasible and efficient strategy to analyze SNP genotypes. Keywords: nested–PCR; single nucleotide polymorphism (SNP); genotyping (single nucleotide polymer- SNP phismSNP)