PCR-CTPP 一种基于错配技术的SNP分型方法的改良精选.pdfVIP

HEREDITAS (Beijing) 2011 2 , 33(2): 182 ―188 ISSN 0253-9772 技术与方法 DOI: 10.3724/SP.J.1005.2011.00182 PCR-CTPP: 一种基于错配技术的SNP分型方法的改良 2, 3 3, 4 1, 2 1, 2 1, 2 1, 2 1, 2 1, 2 王柯 , 章金涛 , 运玉霞 , 吴晓冰 , 陈阿群 , 王鹏 , 王凯娟 , 张建营 , 代丽萍 1, 2 1. , 450001; 2. , 450052; 3. , 450001; 4. , 450052 为探讨两对交叉引物PCR (PCR-CTPP)技术的原理并提高SNP 分型的准确性, 以人类MTHFR 基因 1298 位点突变为例, 通过设计合理的引物、优化引物终浓度及复性温度等重建 PCR-CTPP 检测系统, 对比常规 PCR-CTPP 和改良的PCR-CTPP 检测系统的可靠性。结果表明, 改良的PCR-CTPP 检测系统更加准确, 支持了 常规方法存在理论缺陷的观点; 批量鉴定结果进一步验证了改良方法的可靠性。该技术有望在医学和分子生物 学等领域广泛应用。 两对交叉引物PCR; 四引物扩增受阻突变体系PCR; SNP分型; 人为错配; 亚甲基四氢叶酸还原酶 Improvement on PCR-CTPP: a SNP genotyping approach based on mismatch technique WANG Ke2, 3 , ZHANG Jin-Tao3, 4, YUN Yu-Xia1, 2, WU Xiao-Bing1, 2, CHEN A-Qun1, 2, WANG Peng1, 2, WANG Kai-Juan1, 2, ZHANG Jian-Ying1, 2, DAI Li-Ping1, 2 1. Department of Epidemiology and Biostatistics, College of Public Health, Zhengzhou University, Zhengzhou 450001, China; 2. Henan Key Laboratory of Tumor Epidemiology, Zhengzhou 450052, China; 3. Department of Bioengineering, Zhengzhou University, Zhengzhou 450001, China; 4. Laboratory Animal Center of Zhengzhou University, Zhengzhou 450052, China Abstract: To explore the technique principle of PCR with confronting two-pair primers (PCR-CTPP) and improve the accuracy of SNP genotyping by taking the 1298 locus of human gene MTHFR as an example, the reliability between con- ventional PCR-CTPP and improved PCR-CTPP was compared using reconstructed PCR-CTPP detecting system in terms of designing appropriate primers and optimizing annealing tempera