title: Detecting the polymorphism of TERF1 gene by an improved PCR-RFLP method.
abstract: BACKGROUND
  Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) is a common and mature method of detecting the single nucleotide polymorphism (SNP). But, for the polymorphism site rs3863242 of telomeric repeat binding factor 1(TERF1) gene, there is no appropriate restriction enzyme to recognize it, which limits the research between the variants of rs3863242 and human diseases.
  
  
  METHODS
  The reverse primer was designed based on turning the 3rd base T into the mismatch base G. After PCR amplification, a new restriction enzyme site was introduced into the TERF1 gene amplification products. Two hundred forty samples from Chinese Han individuals were genotyped to evaluate this method.
  
  
  RESULTS
  A new restriction enzyme site for CviQI was introduced into the PCR products. The genotype frequencies of 240 samples from Chinese Han individuals were 4.17% for A/A, 29.58% for A/G, 66.25% for G/G respectively. The allele frequencies were 18.96% for A and 81.04% for G respectively. The genotyping results of PCR products were consistent with the gene sequencing result.
  
  
  CONCLUSIONS
  We developed a simple, direct and economical technique for analyzing the polymorphism of TERF1 rs3863242. It may be applied to the colony screening of other SNPs, mutation-screening of tumor-related gene or mutations in some specific genes on a large scale, in the future.
authors.affiliation: Department of Occupational Health Occupational Disease, College of Public Health, Zhengzhou University, Zhengzhou, China.
authors.forename: Pengpeng
authors.identifier: http://orcid.org/0000-0001-5867-6458
authors.initials: P
authors.lastname: Wang
authors.affiliation: Department of Epidemiology and Biostatistics, College of Public Health, Zhengzhou University, Zhengzhou, China.
authors.forename: Yongli
authors.initials: Y
authors.lastname: Yang
authors.affiliation: Henan Provincial Institute of Occupational Health, Zhengzhou, China.
authors.forename: Sihua
authors.initials: S
authors.lastname: Wang
authors.affiliation: Department of Occupational Health Occupational Disease, College of Public Health, Zhengzhou University, Zhengzhou, China.
authors.forename: Xiaoran
authors.identifier: http://orcid.org/0000-0001-8533-7750
authors.initials: X
authors.lastname: Duan
authors.affiliation: Department of Occupational Health Occupational Disease, College of Public Health, Zhengzhou University, Zhengzhou, China.
authors.forename: Tuanwei
authors.initials: T
authors.lastname: Wang
authors.affiliation: Department of Occupational Health Occupational Disease, College of Public Health, Zhengzhou University, Zhengzhou, China.
authors.forename: Xiaolei
authors.initials: X
authors.lastname: Feng
authors.affiliation: Department of Occupational Health Occupational Disease, College of Public Health, Zhengzhou University, Zhengzhou, China.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Yan
authors.affiliation: Department of Toxicology, College of Public Health, Zhengzhou University Zhengzhou, China.
authors.forename: Yongjun
authors.initials: Y
authors.lastname: Wu
authors.affiliation: Department of Toxicology, College of Public Health, Zhengzhou University Zhengzhou, China.
authors.forename: Songcheng
authors.initials: S
authors.lastname: Yu
authors.affiliation: Department of Occupational Health Occupational Disease, College of Public Health, Zhengzhou University, Zhengzhou, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
mesh_terms: D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length / Q000235:genetics*; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D000089804:Shelterin Complex; D034501:Telomere-Binding Proteins / Q000235:genetics*
chemical_list: D000089804:Shelterin Complex; C059329:TERF1 protein, human; D034501:Telomere-Binding Proteins
keywords: 
  TERF1
  ; PCR-RFLP; Single nucleotide polymorphism; rs3863242