Method for selecting molecular marker for indicating and identifying sheep wool fiber diameters

A technology of sheep wool fiber and molecular markers, which is applied in the field of molecular markers to detect the diameter of sheep wool fibers, achieving the effects of high accuracy, accelerated breeding process, and simple operation

Inactive Publication Date: 2012-02-29
XINJIANG ACADEMY OF AGRI & RECLAMATION SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Recently, multiple SNP sites were found in the coding regions of KAP1.1 and KAP1.3 genes, but

Method used

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  • Method for selecting molecular marker for indicating and identifying sheep wool fiber diameters
  • Method for selecting molecular marker for indicating and identifying sheep wool fiber diameters
  • Method for selecting molecular marker for indicating and identifying sheep wool fiber diameters

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 : The following examples describe the present invention in more detail:

[0037] 1. Experimental materials

[0038] 1. Sample collection and character determination

[0039] Use a disposable syringe to draw about 1ml of blood from the jugular vein of the sheep, and inject it into a 1.5ml centrifuge tube that is autoclaved and contains about 200μl of 2% sterile EDTA (Ethylene diamine tetraacetic acid, EDTA) anticoagulant. Shake gently Evenly, record the sheep number and store at -20℃ for later use.

[0040] Collect the skin hair samples on the side of the sheep. According to the national fiber inspection standards and refer to the International Wool Organization (IWTO) fiber testing standards, the side hair samples of the sheep are measured for fineness, curl, fineness dispersion, natural length, and amount of dirty hair. , Density and net gross rate determination.

[0041] 2. Drugs and enzymes

[0042] Tris (Tris), Sigma Chemicals Co; Tris saturated phenol, Beijing Di...

Embodiment 2

[0118] Reference figure 1 — image 3 : Compared with Example 1, the difference of this example lies in the small amount of sheep genomic DNA extraction:

[0119] (1) Add 20μl of whole blood into a 1.5ml centrifuge tube containing 700μl of 1×SET, and mix gently.

[0120] (2) Add proteinase K (10mg / ml) to a final concentration of 100-200 μg / μl and 10% SDS to a final concentration of 0.5%, and digest at 55°C for 12 hours.

[0121] (3) After the digestion is complete, add an equal volume of Tris saturated phenol, invert back and forth to mix.

[0122] (4) Centrifuge at 12,000 rpm for 10 minutes, carefully move the upper aqueous phase into another centrifuge tube with a tip with a cut tip, and discard the organic phase. Repeat the third and fourth steps once.

[0123] (5) Add an equal volume of phenol, chloroform, and isoamyl alcohol mixture (volume ratio 24:23:1) to the water phase, and mix for 10 minutes. Centrifuge at 12,000 rpm for 10 minutes, and remove the water phase to another cen...

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Abstract

The invention discloses a method for selecting molecular markers for indicating and identifying sheep wool fiber diameters. The method comprises the following steps of: designing a pair of primers according to a gene sequence KAP 1.1 of sheep; extracting deoxyribonucleic acid (DNA) of a genome from sheep blood, and performing polymerase chain reaction (PCR) amplification on the DNA of the genome of the sheep; performing electrophoretic separation on a PCR product by using polyacrylamide gel, and detecting variant loci inserted/deleted by 60 bp and 30 bp in KAP 1.1 gene exons; and performing polymorphic analysis. The method is easy to operate, low in cost and high in accuracy, and can be used for automated detection. By the method, a simple, effective and convenient molecular marker process is provided for the auxiliary selection of the markers in the sheep breeding process, and an effective molecular marker breeding means is provided for the fineness characters of the sheep, so that the breeding process of high-quality fine-wool sheep can be accelerated.

Description

technical field [0001] The invention belongs to the field of animal molecular genetics, in particular to a method for detecting the fiber diameter of sheep wool with molecular markers. Background technique [0002] my country is a large agricultural country, and animal husbandry is an important part of our country's agriculture, and animal husbandry occupies a pivotal position in the national economy. According to statistics, in 2010, the growth rate of animal husbandry income has surpassed that of planting, and it has become one of the main sources of farmers' income. Vigorously developing animal husbandry is still the focus of my country's agricultural industrial structure adjustment, agricultural and rural economic development and farmers' income increase. [0003] Sheep raising is a traditional industry of my country's animal husbandry. Fine-wool sheep occupy an important position in my country's sheep raising industry. Fine-wool sheep are the main animal species of XPC...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 杨华杨永林刘守仁冯静
Owner XINJIANG ACADEMY OF AGRI & RECLAMATION SCI
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