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Method for quickly detecting RFLP (restriction fragment length polymorphism) of SNP (single nucleotide polymorphism) of common ox SIRT1 (silent information regulator 1) gene

A cattle, gene technology, applied in the field belonging to the field of molecular genetics

Inactive Publication Date: 2013-09-11
NORTHWEST A & F UNIV
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Problems solved by technology

[0005] At present, there is no research on the genetic variation of the bovine SIRT1 gene at home and abroad. Since the function of the SIRT1 gene involves important growth traits such as fat metabolism and muscle differentiation, the detection method provided by the present invention lays the foundation for the establishment of the relationship between the SNP of the SIRT1 gene and the growth traits of Chinese yellow cattle. foundation for marker-assisted selection of growth traits in Chinese cattle, and rapid establishment of cattle populations with excellent genetic resources

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  • Method for quickly detecting RFLP (restriction fragment length polymorphism) of SNP (single nucleotide polymorphism) of common ox SIRT1 (silent information regulator 1) gene
  • Method for quickly detecting RFLP (restriction fragment length polymorphism) of SNP (single nucleotide polymorphism) of common ox SIRT1 (silent information regulator 1) gene
  • Method for quickly detecting RFLP (restriction fragment length polymorphism) of SNP (single nucleotide polymorphism) of common ox SIRT1 (silent information regulator 1) gene

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Embodiment Construction

[0024] The present invention uses the PCR-RFLP method to detect the single nucleotide polymorphism at position -274 of the cattle SIRT21 gene, and the present invention will be further described in detail below.

[0025] 1. Collection of Cattle Blood Samples

[0026] The present invention uses the population of 5 local yellow cattle breeds in China as the detection object, and the specific collection samples are shown in Table 1:

[0027] Table 1 Collection of Cattle Samples

[0028]

[0029]

[0030] 2. Separation, extraction and purification of genomic DNA from blood samples

[0031] 1) Thaw the frozen blood sample (mainly blood cells) at room temperature, transfer 500μL to a 1.5mL Eppendorf centrifuge tube, add an equal volume of PBS solution, mix well, centrifuge at 12000r / min for 10min (4℃), discard the supernatant, and repeat the above steps Until the supernatant is transparent and the precipitate is pale yellow;

[0032] 2) Add 500 μL of DNA extraction buffer i...

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Abstract

The invention discloses a method for quickly detecting RFLP (restriction fragment length polymorphism) of SNP (single nucleotide polymorphism) of a common ox SIRT1 (silent information regulator 1) gene promotor region. The method comprises the following steps of: taking whole genome DNA (deoxyribonucleic acid) of common ox blood as a template and a primer pair P (F and R) as a primer; performing PCR (polymerase chain reaction) amplification on the common ox SIRT1 gene promotor region; digesting a PCR amplification product by using restriction endonuclease SmaI; performing agarose gel electrophoresis on segments after enzyme digestion; and identifying the SNP of the common ox SIRT1 promotor region according to a result of the agarose gel electrophoresis. Because important functions including muscle differentiation, lipogenesis and the like related to the SIRT1 are closely related to common ox meat quality characteristics, and the SNP can significantly affect the promotor activity of the SIRT1 gene and is related to various important economic characteristics of the common ox, the detection method provided by the invention lays a foundation for the establishment of the relationship between the SNP and growth characteristics of the SIRT1 gene, and can be used for marker assisted selection of the growth characteristics of the Chinese common ox so as to facilitate rapid establishment of common ox populations with excellent genetic resources.

Description

technical field [0001] The invention belongs to the field of molecular genetics, in particular to a RFLP (restriction fragment length polymorphism) method for rapidly detecting the single nucleotide polymorphism (SNP) of the silence regulator 1 (SIRT1) gene, specifically, A method of digesting the gene sequence containing the single nucleotide polymorphism site by using a restriction endonuclease, separating the fragment size according to agarose gel electrophoresis, and analyzing the fragment size by using a gel imaging system, To determine its SNP. Background technique [0002] Single nucleotide polymorphism (SNP) refers to the polymorphism caused by the substitution of a single nucleotide (A / T / C / G) in the genomic DNA sequence. SNP belongs to the third-generation molecular marker, which has the advantages of high density, biallelic genes, and easy automatic detection. Because SNP has the advantages of other markers, it has been widely used in various fields of life scienc...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 陈宏李明勋孙晓梅蓝贤勇
Owner NORTHWEST A & F UNIV
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