Single Nucleotide Polymorphism Site of Cattle wnt10b Gene and Its Detection Method

A single nucleotide polymorphism and detection method technology, applied in the field of molecular genetics, can solve the problems of cumbersome operation and high cost

Inactive Publication Date: 2016-09-14
XUZHOU NORMAL UNIVERSITY
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Problems solved by technology

The RFLP-PCR method not only has the accuracy of DNA sequencing, but also overcomes the shortcomings of expensive, cumbersome operations, and false positives, and there are no special requirements for the sequence sites detected

Method used

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  • Single Nucleotide Polymorphism Site of Cattle wnt10b Gene and Its Detection Method
  • Single Nucleotide Polymorphism Site of Cattle wnt10b Gene and Its Detection Method
  • Single Nucleotide Polymorphism Site of Cattle wnt10b Gene and Its Detection Method

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Embodiment Construction

[0047] In order to facilitate the understanding of the technical solution of the present invention, the present invention will be further described below in conjunction with specific examples.

[0048] The following is through the collection of cattle samples and genomic DNA extraction, detection, purification and concentration analysis, the PCR amplification of the first intron and the second, fourth and fifth exons of the cattle WNT10B gene, the first intron of the cattle WNT10B gene and the The PCR-RFLP analysis embodiment of the 2nd, 4th, 5th exon will further illustrate the technology of the present invention and its effect. What has been described is by way of explanation, not limitation, of the invention.

[0049] 1. Amplification of partial DNA sequence of cattle WNT10B gene and detection of its polymorphism

[0050] 1. Collection and processing of cattle blood samples

[0051] Take 10 mL of cattle blood sample, add 500 μL of 0.5 mol / L EDTA to anticoagulate, slowly i...

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Abstract

The invention discloses a method for rapidly detecting the single nucleotide polymorphism of the cattle WNT10B gene. The whole genome DNA of the cattle to be tested including the WNT10B gene is used as a template, and the primer pair P1 and P3 are used as primers to amplify the cattle WNT10B gene by PCR. , with restriction endonucleases NaeI and ApaI to digest the PCR products amplified by primer pairs P1 and P3 respectively, and then carry out agarose gel electrophoresis to the amplified fragments after enzyme digestion; identify yellow cattle according to the results of agarose gel electrophoresis Single nucleotide polymorphisms at positions 220 and 3980 of the WNT10B gene. The method of the invention is a method for screening and detecting molecular genetic markers closely related to the growth and development traits of cattle at the DNA level, so as to be used for auxiliary selection and molecular breeding of cattle, and to accelerate the breeding speed of fine cattle breeds.

Description

technical field [0001] The invention belongs to the field of molecular genetics, relates to the detection of gene single nucleotide polymorphism (SNP), in particular to a single nucleotide polymorphism for detecting intron 1 and exon 2, 4, 5 of cattle WNT10B gene. state-of-the-art detection method. 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. Therefore, commonly referred to as SNPs include base substitutions, insertions, deletions, and changes in the copy number of repeated sequences. A SNP indicates that there is a nucleotide change at a certain position in the genome, which is mainly caused by the conversion or transversion of a single base; SNPs with conversion mutations account for about 2 / 3, and other SNPs are in similar level. The cytosine of the CpG dinucleotide is the most mutated site in the genome, most of which are methylated...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/12C12Q1/68
Inventor 陈宏赵静张春雷房兴堂
Owner XUZHOU NORMAL UNIVERSITY
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