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PCR-RFLP method for detecting single nucleotide polymorphism of gene CREB 1

A technique of PCR-RFLP and single nucleotide polymorphism, which is applied in the fields of biochemical equipment and methods, measurement/inspection of microorganisms, etc., can solve the problems that the correlation of CREB1 gene type II diabetes has not been reported yet, and is easy to be popularized and applied , low cost, high precision effect

Inactive Publication Date: 2020-01-31
WUHAN UNIV OF SCI & TECH
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Problems solved by technology

[0007] At present, the research on CREB1 gene at home and abroad is mainly focused on its function and regulatory mechanism, and there is no report on the genetic variation of CREB1 gene and its correlation with type 2 diabetes.

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  • PCR-RFLP method for detecting single nucleotide polymorphism of gene CREB 1
  • PCR-RFLP method for detecting single nucleotide polymorphism of gene CREB 1
  • PCR-RFLP method for detecting single nucleotide polymorphism of gene CREB 1

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Embodiment Construction

[0027] The present invention will be further described in detail below, which is an explanation of the present invention rather than a limitation. Unless otherwise specified, the routine experimental conditions or the conditions suggested by the manufacturer's instruction were followed.

[0028] The PCR-RFLP method that is used to detect gene CREB1 single nucleotide polymorphism is characterized in that: take the blood genome DNA of diabetic patient and normal person as template, take primer pair P (F, R) as primer, PCR amplification CREB1 gene, and then use restriction endonuclease to digest it, and then electrophoresis detection can accurately identify the single nucleotide polymorphism of the sample to be tested; that is, in the presence of Taq DNA polymerase, buffer environment, Mg++, dNTPs In the case of P, use polymerase chain reaction primers to amplify P by PCR, then use restriction endonuclease to digest it, and then detect by electrophoresis to accurately identify th...

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Abstract

The invention discloses a PCR-RFLP method for detecting the single nucleotide polymorphism of a gene CREB 1. Firstly, according to a DNA pool sequencing result, the detected gene polymorphism comprises single nucleotide polymorphism of T or G at the 1354th site from a CREB1 gene promoter region to a transcription starting point and single nucleotide polymorphism of T or A at the 1343th site. PCR amplification is conducted on a CREB1 gene sequence containing polymorphic sites in the presence of Taq DNA polymerase, Buffer (a buffer environment), Mg++ and dNTPs by taking genome DNA of a to-be-detected sample as a template and a primer pair P as a primer, a PCR product is divided into two parts, the two parts are digested by using restriction enzymes Hha I and Xsp I respectively, and enzyme digestion products are subjected to typing through agarose gel electrophoresis.

Description

technical field [0001] The invention belongs to the technical field of molecular biology detection, and in particular relates to a PCR-RFLP method for detecting single nucleotide polymorphism of gene CREB1. Background technique [0002] Diabetes is considered to be the fifth most serious disease that endangers human health in the world, and 90% of them are non-insulin-dependent diabetes, also known as type II diabetes. The disease is a complex metabolic disorder, and its pathogenesis is complex, which is the result of the interaction between genetics and the environment, among which genetic polymorphism may be the main factor affecting the susceptibility of the disease among individuals. Since the early clinical symptoms of type II diabetes are not obvious, patients often delay the best time for treatment. Therefore, finding and identifying genetic markers related to type 2 diabetes in the human genome will be beneficial to the early screening and diagnosis of the disease, ...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12Q1/683
CPCC12Q1/683C12Q1/6883C12Q2600/156C12Q2531/113
Inventor 徐瑶宋如晦石伟林代洋许娜廖兴华张同存
Owner WUHAN UNIV OF SCI & TECH
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