Carcinogenesis, Teratogenesis & Mutagenesis ›› 2022, Vol. 34 ›› Issue (2): 129-133.doi: 10.3969/j.issn.1004-616x.2022.02.010

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Development of mismatched long PCR primers for analysis of rs3093816 polymorphism in the CCNH gene

CHANG Wei1, DING Mingcui2, WANG Wei2   

  1. 1. Center for Disease Control, General Hospital of Pingmei Shenma Medical Group, Pingdingshan 467002;
    2. Department of Occupational Health and Occupational Disease, College of Public Health, Zhengzhou University, Zhengzhou 450001, Henan, China
  • Received:2021-09-06 Revised:2022-02-22 Published:2022-04-07

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Abstract: OBJECTIVE：It has been demonstrated that the single nucleotide polymorphism (SNP) in the rs3093816 site of the cyclin H (CCNH) gene was associated with an increased cancer risk. There is，however， a limitation to utilizing PCR-RFLP due to the lack of proper restriction enzyme sites in rs3093816. Therefore， an appropriate method for identifying the polymorphism was developed in this study. METHODS： A new restriction site was introduced into the CCNH amplification products using a created-restriction site PCR (CRSPCR) which allowed the enzymes CviQ I in distinguishing the PCR products. In addition，long primers were compared with relatively short primers to investigate whether the long primers are more economical and modest. RESULTS： Compared with the short primers， the larger the proportion of the long primers to the total amplified fragment， the easier to distinguish the digestion products， and the shorter the time required for electrophoresis. CONCLUSION：A mismatched long PCR primers method was successfully developed to detect the rs3093816 polymorphism in the CCNH gene.

Key words: cyclin H, created restriction site, single nucleotide polymorphism, PCR-RFLP

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