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

• en • Previous Articles    

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. 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

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