Detection of the rs10250202 polymorphism in protection of telomeres 1 gene through introducing a new restriction enzyme site for PCR–RFLP assay

Abstract

Human protection of telomeres 1 (POT1) gene is a single stranded telomere binding proteins with a critical role in ensuring chromosome stability. There have been variants of POT1 gene, and the polymorphisms of POT1 gene were associated with some diseases. Polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) is a traditional method to detect the single nucleotide polymorphism (SNP), and it can be used to detect the polymorphism of rs10250202. But the restriction enzymes required for the detection of the polymorphism of rs10250202 are expensive. So we designed a novel PCR–RFLP assay for genotyping the POT1 rs10250202 SNP. In the study, a new restriction enzyme cutting site was created by created restriction site PCR (CRS-PCR), and the restriction enzyme BclI for CRS-PCR was cheaper than other enzymes. After detecting Han Chinese workers, Allele frequencies were found to be 51.54 % for allele A and 48.46 % for allele C respectively. The PCR results were confirmed by DNA sequencing. CRS-PCR provides a simple, low-cost, practical, and reproducible method.

Site-Specific Sequence Exchange Between Homologous and Non-homologous Chromosomes

“…Restriction endonuclease analysis was carried out to distinguish between TS537 and TS * 537 * . However, because the CRISPRmediated changes in TS * 537 * did not destroy an existing restriction site, CRS-PCR (created restriction site PCR) (Qiao et al, 2013;Wang et al, 2016;Avanus and Altınel, 2017;Ding et al, 2017) and overlapping PCR were used to create a restriction site for the TS537 PCR product, but not for the TS * 537 * PCR product. Specific steps are outlined in Figure 2H as follows: primers h + e, h + f, e + k, and f + k can only be amplified from F1 + cre; one or two bases were changed near the 3 -end of the oligonucleotide m or o, respectively (Supplementary Figure 1); primers h + n and m + i created the overlapping PCR product hi, and similarly, primers j + p and o + k created the overlapping PCR product j-k.…”

Mapping Growth Hormone Gene of Body Weight Krui Cattle in Pesisir Barat Regency Lampung, Indonesia

“…If the relationship can be found and the relationship is close enough and is characteristic of the entire population, then it can be used for selection as an indicator of productivity. One way to detect polymorphisms is through the PCR-RFLP method 8 .…”