NDLI: Application of created restriction site PCR-RFLP to identify POT1 gene polymorphism

Content Provider	Springer Nature Link
Author	Wang, Tuanwei Wang, Sihua Duan, Xiaoran Feng, Xiaolei Wang, Pengpeng Yao, Wu Wu, Yongjun Feng, Feifei Yu, Songcheng Wu, Yiming Wang, Wei
Copyright Year	2016
Abstract	Protection of telomeres protein 1 (POT1) plays pivotal roles in protection of chromosome ends and regulation of telomere length with other telomere binding proteins; its genetic polymorphisms are associated with many diseases. In this study, we explored a novel PCR-RFLP method for typing the single nucleotide polymorphism (SNP) rs1034794 of the human POT1 gene. A new restriction enzyme site was introduced into a POT1 gene amplification product by created restriction site PCR (CRS-PCR). One primer was designed based on changed sequence; after PCR amplification, a new restriction enzyme site for AluI was introduced into the PCR products. One hundred and seventy eight samples from Han Chinese individuals were tested to evaluate this new method. The 3′-end of the forward primer was next to the polymorphic site, and the third base from the 3′-end was the mismatched base A. The final PCR product contained the AGCT sequence (AluI recognition site) when the ancestral POT1 alleles were amplified. The data obtained with the new method perfectly matched those obtained with the sequencing method. Thus, CRS-PCR is a new low-cost and high-efficiency alternative for rs1034794 typing.
Starting Page	624
Ending Page	627
Page Count	4
File Format	PDF
ISSN	00062979
Journal	Biochemistry (Moscow)
Volume Number	81
Issue Number	6
e-ISSN	16083040
Language	English
Publisher	Pleiades Publishing
Publisher Date	2016-06-14
Publisher Place	Moscow
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	created restriction site single nucleotide polymorphism protection of telomeres 1 Biochemistry Bioorganic Chemistry Microbiology Biomedicine general
Content Type	Text
Resource Type	Article
Subject	Biochemistry Biophysics Geriatrics and Gerontology

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4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
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