Pengpeng Wang1, Yongli Yang2, Sihua Wang3, Xiaoran Duan1, Tuanwei Wang1, Xiaolei Feng1, Zhen Yan1, Yongjun Wu4, Songcheng Yu4, Wei Wang1. 1. Department of Occupational Health Occupational Disease, College of Public Health, Zhengzhou University, Zhengzhou, China. 2. Department of Epidemiology and Biostatistics, College of Public Health, Zhengzhou University, Zhengzhou, China. 3. Henan Provincial Institute of Occupational Health, Zhengzhou, China. 4. Department of Toxicology, College of Public Health, Zhengzhou University Zhengzhou, China.
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.
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 TERF1rs3863242. 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: Myriam Brossard; Shenying Fang; Christopher I Amos; Florence Demenais; Amaury Vaysse; Qingyi Wei; Wei V Chen; Hamida Mohamdi; Eve Maubec; Nolwenn Lavielle; Pilar Galan; Mark Lathrop; Marie-Françoise Avril; Jeffrey E Lee Journal: Int J Cancer Date: 2015-05-26 Impact factor: 7.396
Authors: Diana Suárez-Salgado; Gaspar Manuel Parra-Bracamonte; Flaviano Benavides-González; Isidro O Montelongo Alfaro; Ana María Sifuentes Rincón; Victor Ricardo Moreno-Medina; Xochitl Fabiola De la Rosa-Reyna Journal: Mol Biol Rep Date: 2019-10-01 Impact factor: 2.316