Molecular marking method for predicting and identifying length of sheep wool

A molecular marker, sheep technology, applied in the direction of biochemical equipment and methods, microbial determination/inspection, etc., to achieve the effect of accelerating the breeding process, high accuracy, and simple operation

CN103276098BInactive Publication Date: 2014-07-16NORTHEAST AGRICULTURAL UNIVERSITY
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
Publication Date
2014-07-16
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention relates to a molecular marking method, in particular to a molecular marking method for predicting and identifying the length of sheep wool. The method comprises the steps as follows: 1), extracting sheep genome DNA, designing a primer, performing PCR (polymerase chain reaction) amplification, performing enzyme digestion, and obtaining an enzyme digestion product; 2), subjecting the enzyme digestion product to electrophoretic separation, and judging genotypes according to an electrophoretic separation result; 3), performing association analysis and estimating the least square mean value of characters, and obtaining a result that the length of wool of a GG genotype individual is remarkably higher than that of wool of AA genotype and AG genotype individuals in the three genotypes; and 4), constructing wool length character breeding population taking the GG genotype individual as the primary so that the length of the sheep wool is predicated and identified. According to the method, the operation is simple, the cost is low, the accuracy is high, and automatic detection can be performed. Selective breeding of the sheep wool length is performed with the molecular marking method, so that a genetic breeding process of the length characters of the sheep wool can be accelerated, breeding sheep can be early selected, the sheep can be selected and left after birth, and a sheep breeding process is accelerated.
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Description

technical field

[0001] The invention relates to a molecular marking method. Background technique

[0002] Fine-wool sheep occupy an important position in my country's animal husbandry industry, and the main product of fine-wool sheep is wool. As an important textile raw material, fine wool has high economic value. The production of fine-wool sheep is not only related to the economic development and social stability of the producing areas, but also to the development of my country's wool spinning industry and the balance of import and export trade. With the increasing demand for wool at home and abroad, the cultivation of high-quality fine-wool sheep has become an urgent problem to be solved in the field of wool production and sheep breeding.

[0003] The breeding of Chinese Merino sheep (Xinjiang military reclamation type) began in 1972, and has successively bred six strains, namely military reclamation type A strain, military reclamation type B strain, ultra-fine strain, ...

Examples

specific Embodiment approach 1

[0021] Embodiment 1: In this embodiment, the molecular marker method for predicting and identifying the length of sheep wool is carried out according to the following steps:

[0022] 1. Using the phenol / chloroform method to extract sheep genomic DNA from the ear tissue of Chinese Merino sheep, design primers DKK1F1 and DKK1R1 according to the c.576A>G site of the fourth exon region of the sheep DKK1 gene, and then analyze the sheep genome Carrying out PCR amplification of the DNA to obtain a PCR amplification product, and then digesting the PCR amplification product with endonuclease Pvu I to obtain a digestion product;

[0023] 2. Use agarose gel with a concentration of 2.5% to electrophoresis separate the digested products, and then determine the genotype according to the electrophoresis separation results. The criteria for determination: ① Electrophoresis presents a band with a size of 119bp, which means that the sheep DKK1 gene is 4th The c.576A>G site in the exon region w...

specific Embodiment approach 2

[0054] Specific embodiment two: the difference between this embodiment and specific embodiment one is that the reaction system for PCR amplification in step one is a 10 μL reaction system, which consists of the following components:

[0055]

[0056] PCR amplification conditions were: 94°C pre-denaturation for 5 min, 94°C denaturation for 30 s, 52°C annealing for 30 s, 72°C extension for 8 s, a total of 33 cycles, 72°C extension for 7 min, and 4°C incubation. Others are the same as in the first embodiment.

specific Embodiment approach 3

[0057] Specific embodiment three: the difference between this embodiment and specific embodiment one or two is that the enzyme digestion system in step one is as follows:

[0058]

[0059] Enzyme digestion conditions are: 37 ° C enzyme digestion 1-2h. Others are the same as in the first or second embodiment.

[0060]