Method for detecting single nucleotide polymorphism of cattle krupple-like factor (KLF) 7 gene

A single nucleotide polymorphism and detection method technology, which is applied in the direction of microbial measurement/testing, biochemical equipment and methods, and can solve problems such as lack of research

Inactive Publication Date: 2010-12-01
NORTHWEST A & F UNIV
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Problems solved by technology

Due to the current lack of research in the field of genetic variation of the KLF7 gene in Chinese yellow cattle, the research on the function of this gene locus and the relationship between the genetic variation of this gene and growth and development traits, as well as the application of it to molecular genetic markers and molecular breeding are still in a blank state

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  • Method for detecting single nucleotide polymorphism of cattle krupple-like factor (KLF) 7 gene
  • Method for detecting single nucleotide polymorphism of cattle krupple-like factor (KLF) 7 gene
  • Method for detecting single nucleotide polymorphism of cattle krupple-like factor (KLF) 7 gene

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Embodiment Construction

[0043] The following is through the collection of cattle samples and genomic DNA extraction, detection, purification and concentration analysis, PCR amplification of exon 2 of cattle KLF7 gene and its flanking region, TaqI CRS of DNA fragments of cattle KLF7 gene exon 2 and its flanking region - PCR-RFLP analysis embodiment to further illustrate the technology of the present invention and its effects. What has been described is by way of explanation, not limitation, of the invention.

[0044] a. Cloning of partial DNA sequence of cattle KLF7 gene and detection of its polymorphism

[0045] 1. Collection and processing of cattle blood samples

[0046] Take 10 mL of cattle blood sample, 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.

[0047] The present invention has adopted a total of 1002 unrelated cow blood samples from 4 cattle varieties, specifically:

[0048] (1) Nanyang cattle blood sample...

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Abstract

The invention discloses a method for fast detecting the single nucleotide polymorphism (SNP) of a cattle krupple-like factor (KLF) 7 gene. The method comprises the following steps of: performing polymerase chain reaction (PCR) amplification on the cattle KLF7 gene by taking a whole genome DNA comprising a KLF7 gene of cattle to be detected as a template and the mixture of primer pairs P1, P2, P3 and the like in a mol ratio as a primer; after digesting a PCR amplification product by using a restriction enzyme TaqI, performing agarose gel electrophoresis on an amplified fragment subjected to enzyme digestion; and according to the result of the agarose gel electrophoresis, identifying the SNP of the 41041st, 42025th and 42075th positions of the cattle KLF7 gene. In the method, the SNPs of the KLF7 gene are subjected to genotyping and gene frequency analysis, and the growth traits of Nanyang cattle in different growth stages are subjected to trait association analysis. The results show that the SNP locus for the KLF7 gene, particularly P2, can serve as a molecular marker selected in the early stage (6 month age and 12 month age) of the growth traits of the cattle.

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 method for detecting the single nucleotide polymorphism of cattle KLF7 gene exon 2 and its flanking region . 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 and can be spontane...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 陈宏马亮蓝贤勇李转见王景淮永涛雷初朝胡沈荣
Owner NORTHWEST A & F UNIV
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