Polymerase chain reaction (PCR)-restricted fragment length polymorphism (RFLP) method for quickly detecting single nucleotide polymorphism of goat Lhx3 gene

A PCR-RFLP, single nucleotide polymorphism technology, applied in the field of molecular genetics, can solve the problem that the goat Lhx3 gene polymorphism has not been reported, and achieve the effect of low cost and high precision

Inactive Publication Date: 2010-12-22
NORTHWEST A & F UNIV
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Problems solved by technology

[0006] At present, there is no report on the polymorphism of goat Lhx3 gene and its detection method

Method used

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  • Polymerase chain reaction (PCR)-restricted fragment length polymorphism (RFLP) method for quickly detecting single nucleotide polymorphism of goat Lhx3 gene
  • Polymerase chain reaction (PCR)-restricted fragment length polymorphism (RFLP) method for quickly detecting single nucleotide polymorphism of goat Lhx3 gene
  • Polymerase chain reaction (PCR)-restricted fragment length polymorphism (RFLP) method for quickly detecting single nucleotide polymorphism of goat Lhx3 gene

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Embodiment Construction

[0032] The present invention designs primers according to the conserved sequence of the Lhx3 gene, respectively uses the genomic DNA pools of four goat varieties as templates, performs PCR amplification, purifies the PCR products, and obtains the partial sequence of the Lhx3 gene of goats such as HM446543 after sequencing. The present invention will be described in detail below, which is an explanation of the present invention rather than a limitation.

[0033] I. Cloning and DNA sequencing of partial DNA sequence of goat Lhx3 gene

[0034] 1. Collection and processing of goat blood samples

[0035] Take 10 mL of goat blood, add 500 μL of 0.5 mol / L EDTA to anticoagulate, slowly invert 3 times, put it in an ice box, and store it at -80°C for later use.

[0036] The present invention adopts 4 kinds of goat breeds totaling 1073 individuals, including 2 milk goat breeds, 1 cashmere goat breed and 1 meat goat breed, specifically: (1) 229 blood samples of Guanzhong goat were collec...

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Abstract

The invention discloses a PCR-RFLP method for quickly detecting the single nucleotide polymorphism (SNP) of a goat Lhx3 gene. The genetic polymorphism is base polymorphism of T or C at a position-5557 of the goat Lhx3 gene. The method for detecting the SNP of the goat Lhx3 gene comprises the following steps of: performing PRC amplification on the goat Lhx3 gene by using the DNA of a goat genome to be detected which comprises the Lhx3 gene as a template and using a primer pair P as primers; and digesting a PCR amplification product by using a restriction enzyme, performing agarose gel electrophoresis on digested segments, and determining the SNP on the position-5557 of a goat according to the result of the agarose gel electrophoresis. The method is easy to operate, quick, low in cost, high in detection precision and convenient to popularize and use.

Description

technical field [0001] The invention belongs to the field of molecular genetics, in particular to a PCR-RFLP (PCR-restriction fragment length polymorphism) method for rapidly detecting goat Lhx3 gene single nucleotide polymorphism (SNP). Background technique [0002] Single nucleotide polymorphism (single nucleotide polymorphism, SNP) refers to a kind of biallelic or dimorphic genetic variation caused by a single nucleotide variation in genomic DNA, that is, there are two different base. Nucleotide variations include conversion, transversion, insertion, deletion, etc., and the frequency of at least one allele in the population is not less than 1%. Studies have shown that single-base mutations, especially those in the coding region, can affect individual phenotypes. Therefore, SNP-based molecular marker technology can be used for the breeding of superior breeds of livestock. Since SNPs are di-allelic molecular markers, theoretically in a diploid biological population, SNPs ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 陈宏刘金彪蓝贤勇徐瑶张亚雷初朝
Owner NORTHWEST A & F UNIV
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