PCR-CTPP: 一种基于错配技术的SNP分型方法的改良

doi:10.3724/SP.J.1005.2011.00182

遗传 ›› 2011, Vol. 33 ›› Issue (2): 182-188.doi: 10.3724/SP.J.1005.2011.00182

• 技术与方法 • 上一篇

PCR-CTPP: 一种基于错配技术的SNP分型方法的改良

王柯^{2, 3}, 章金涛^{3, 4}, 运玉霞^{1, 2}, 吴晓冰^{1, 2}, 陈阿群^{1, 2}, 王鹏^{1, 2}, 王凯娟^{1, 2}, 张建营^{1, 2}, 代丽萍^{1, 2}

1. 郑州大学公共卫生学院流行病与卫生统计学系, 郑州 450001;
2. 河南省肿瘤流行病学重点实验室, 郑州 450052;
3. 郑州大学生物工程系, 郑州 450001;
4. 郑州大学实验动物中心, 郑州 450052

收稿日期:2010-08-23 修回日期:2010-11-05 出版日期:2011-02-20 发布日期:2011-02-25
通讯作者: 代丽萍 E-mail:lpdai@zzu.edu.cn
基金资助:
国家自然科学基金项目(编号：30872181, 81072360), 河南省高校科技创新人才支持计划(编号：2010HASTIT027)和河南医学科技攻关项目(编号：200903009)资助

Improvement on PCR-CTPP: a SNP genotyping approach based on mismatch technique

WANG Ke^{2, 3}, ZHANG Jin-Tao^{3, 4}, YUN Yu-Xia^{1, 2}, WU Xiao-Bing^{1, 2}, CHEN A-Qun^{1, 2}, WANG Peng^{1, 2}, WANG Kai-Juan^{1, 2}, ZHANG Jian-Ying^{1, 2}, DAI Li-Ping^{1, 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

Received:2010-08-23 Revised:2010-11-05 Online:2011-02-20 Published:2011-02-25
Contact: DAI Li-Ping E-mail:lpdai@zzu.edu.cn

摘要/Abstract

摘要： 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。

关键词: 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

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 conventional PCR-CTPP and improved PCR-CTPP was compared using reconstructed PCR-CTPP detecting system in terms of designing appropriate primers and optimizing annealing temperature and the final concentration of primers. The improved PCR-CTPP detection system proved to be more accurate, which supported the viewpoint on the theoretical defects of conventional PCR-CTPP. The large-scale study verified the reliability of the improved method. It is expected that this improved technique would be widely used in the field of medicine and molecular biology.

Key words: PCR-CTPP, Tetra-primer ARMS PCR, SNP genotyping, artificial/deliberate mismatch, MTHFR

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. PCR-CTPP: 一种基于错配技术的SNP分型方法的改良[J]. 遗传, 2011, 33(2): 182-188.

WANG Ke, ZHANG Jin-Chao, YUN Yu-Xia, TUN Xiao-Bing, CHEN A-Qun, WANG Feng, WANG Kai-Juan, ZHANG Jian-Ying, DAI Li-Ping. Improvement on PCR-CTPP: a SNP genotyping approach based on mismatch technique[J]. HEREDITAS, 2011, 33(2): 182-188.

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PCR-CTPP: 一种基于错配技术的SNP分型方法的改良

Improvement on PCR-CTPP: a SNP genotyping approach based on mismatch technique

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