Open AccessDetecting the polymorphism of TERF 1 gene by an improved PCR ‐ RFLP method
Author(s) -
Wang Pengpeng,
Yang Yongli,
Wang Sihua,
Duan Xiaoran,
Wang Tuanwei,
Feng Xiaolei,
Yan Zhen,
Wu Yongjun,
Yu Songcheng,
Wang Wei
Publication year - 2018
Publication title -
journal of clinical laboratory analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 50
eISSN - 1098-2825
pISSN - 0887-8013
DOI - 10.1002/jcla.22171
Subject(s) - genotyping , restriction enzyme , restriction fragment length polymorphism , single nucleotide polymorphism , genotype , polymerase chain reaction , biology , microbiology and biotechnology , restriction site , genetics , gene , snp genotyping , primer (cosmetics) , variants of pcr , polymorphism (computer science) , snp , allele , chemistry , organic chemistry
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( TERF 1 ) 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 TERF 1 gene amplification products. Two hundred forty samples from Chinese Han individuals were genotyped to evaluate this method. Results A new restriction enzyme site for Cvi QI 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 TERF 1 rs3863242. It may be applied to the colony screening of other SNP s, mutation‐screening of tumor‐related gene or mutations in some specific genes on a large scale, in the future.