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Method for selecting molecular marker for goat yeaning traits

A technology of molecular markers and goats, which is applied in biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of expensive detection, non-existence, cumbersome operation, etc., to speed up breeding progress, accurately estimate, and improve selection efficiency Effect

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

Among these SNP detection techniques, DNA sequence determination is the most accurate SNP detection method, but its detection cost is extremely expensive, and large-scale instruments such as DNA sequencers are required. At the same time, very skilled technicians and Experience, so DNA sequence determination is not an ideal SNP detection method for actual production; of course, the combination of PCR-SSCP and DNA sequencing can be used to detect SNPs can appropriately reduce the detection cost, but the experimental process of PCR-SSCP is relatively long , the operation is cumbersome, and there are false negative problems in the experimental process, so it is not an ideal SNP detection method; as a new SNP detection method, the AS-PCR method has very broad prospects in the future application field, but , this method needs to design special primers, and it can only target specific gene loci. At the same time, there is a probability of false detection in the detection process. Therefore, it is not generally applicable at present; Ligation reaction technology to detect SNP sites requires detection platforms such as plate reader, gene chip, microsphere array technology and mass spectrometer, which is not very practical for general molecular laboratories

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  • Method for selecting molecular marker for goat yeaning traits
  • Method for selecting molecular marker for goat yeaning traits
  • Method for selecting molecular marker for goat yeaning traits

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Embodiment Construction

[0037] A. PCR amplification of intron 1 and 6 of KITL gene and detection of its polymorphism

[0038] 1. Collection and processing of goat blood samples

[0039] Take 5 mL of goat blood sample, add 0.2 μL of ACD (2.4 g of citric acid; 6.6 g of trisodium citrate; 7.35 g of glucose; dilute to 50 mL, autoclave) for anticoagulation, slowly invert 3 times and put it in an ice box, Store at -80°C for later use.

[0040] In this example, a total of 681 ewe blood samples from 3 goat populations were used, specifically: 290 blood samples of Xinong Saneng sheep, collected from Qianyang Saneng sheep breeding farm in Shaanxi Province; 193 blood samples of Guanzhong dairy goats, collected from Shaanxi Zhouzhi Green New Century Biological Co., Ltd.; 198 Boer goat blood samples were collected from the Boer goat breeding farm in Linyou County, Shaanxi.

[0041] 2. Extraction and purification of genomic DNA from blood samples

[0042] 1) Thaw the frozen blood sample at room temperatur...

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Abstract

The invention discloses a method for selecting a molecular marker for goat yeaning traits, which comprises the following steps of: taking a goat genome DNA sequence as a template, amplifying KITL gene introns 1 and 6 by using primers P1 and P2 under a PCR condition in the presence of TaqDNA polymerase, buffer environment, Mg2+ and dNTPs respectively, and judging the size of a destination fragment according to an agarose gel electrophoresis result; digesting the PCR amplification product of the primer P1 by using restriction enzyme CviAII, and then detecting the enzyme digested amplification fragment by using polyacrylamide gel electrophoresis, wherein the amplification product of the primer P1 has mutation of two basic groups; detecting the PCR amplification product of the primer P2 by adopting the polyacrylamide gel electrophoresis, wherein the amplification product of the primer P2 has mutation of one basic group; and then performing gene analysis and gene frequency analysis on the amplification products of the primers P1 and P2, and performing association analysis between the amplification products and the yeaning numbers of Guanzhong dairy goats, western Saanen dairy goats and Boer goats, wherein the analysis results show that an SNPs site of the KITL gene, detected by the primers P1 and P2, can be taken as the molecular marker for goat yeaning trait selection.

Description

technical field [0001] The invention belongs to the field of molecular genetics, and specifically relates to a molecular marker selection method for lambing traits of goats. The method adopts detection of mutation polymorphisms of goat stem cell factor (Kit ligand, KITL) gene intron 1 and 6 bases, and Analysis of its effect on litter size showed that the SNPs of KITL gene detected by primers P1 and P2 could be used as molecular markers for lambing traits selection in goats. Background technique [0002] With the rapid development of molecular biology technology, especially the advent of Polymerase Chain Reaction (Polymerase Chain Reaction) technology and the improvement of electrophoresis technology, various molecular genetic marker technologies have emerged. Breeding of varieties provides new ways and methods. In particular, the combination of molecular marker technology and conventional breeding technology has given birth to a brand-new selection method - marker-assisted ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 曹斌云安小鹏侯金星王建刚宋宇轩杨明明朱广琴王韵斐崔易虹陈秋菊
Owner NORTHWEST A & F UNIV
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