Screening method of goat COX II gene segment capable of serving as molecular marker and application thereof

A technology of gene fragment and molecular marker, applied in the field of livestock molecular marker preparation

Inactive Publication Date: 2012-07-04
ANHUI AGRICULTURAL UNIVERSITY +1
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  • Abstract
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Problems solved by technology

At present, the research on COX II gene mainly focuses on the regulation of liver, heart and other tissue ce

Method used

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  • Screening method of goat COX II gene segment capable of serving as molecular marker and application thereof
  • Screening method of goat COX II gene segment capable of serving as molecular marker and application thereof
  • Screening method of goat COX II gene segment capable of serving as molecular marker and application thereof

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Embodiment Construction

[0023] figure 1 The operation process of this embodiment is shown, and the present invention will be further described below in conjunction with the embodiment.

[0024] 1. Amplification of partial DNA fragments of COX II gene related to goat sperm motility traits

[0025] 1.1 Sample collection and genomic DNA extraction

[0026] Sperm collection is performed by the pseudo-vaginal method. The rams collected for semen were strong and good in reproductive performance Anhui white goats (40) and Boer goats (40). The samples were provided by the breeding farm of Hefei Boda Animal Husbandry Technology Company. Store frozen in a -70°C freezer. Genomic DNA was extracted according to the following methods:

[0027] 1) Take 1ml of goat semen and put it into a 2ml high-pressure sterilized centrifuge tube, add 1ml of PBS to rinse, discard the supernatant, then add 1ml of PBS to rinse once, and discard the supernatant.

[0028] 2) Add 700 μl of TE extract to the centrifuge tube, then ...

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Abstract

The invention discloses a screening method of a goat COX II gene segment capable of serving as a molecular marker and application thereof. The screening method comprises the following steps: by using constructed goat sperm pool DNA (deoxyribonucleic acid) as a template, using a designed specific primer P1 set a corresponding program in the presence of Tag DNA polymerase, buffer, Mg<2+> and dNTPs (deoxyribonucleotide triphosphates), and carrying out PCR (polymerase chain reaction) amplification on the COX II gene; determining the size of the target segment according to an agarose gel electrophoretogram; sequencing to obtain sequence and mutant site information, wherein the result indicates that the amplification product has one base mutant; after digesting HindI II the P1 amplification product with restriction endonuclease, detecting the digested amplification segment by agarose gel electrophoresis; and analyzing the genotype and gene frequency of the P1 amplification product as well as the correlation of the sperm activities between the Anhwei white goat and Bohr goat, wherein the result indicates that the SNP (single nucleotide polymorphism) site of the COX II gene detected by the primer P1 can serve as a molecular marker for selecting goat sperm activity property, thereby providing a molecular marker for auxiliary selection of a goat sperm quality property marker.

Description

technical field [0001] The invention belongs to the technical field of livestock molecular marker preparation, in particular to a screening method and application of a goat COX II gene fragment which can be used as a molecular marker. Background technique [0002] Sheep semen freezing and artificial insemination is a mature biotechnology that has been put into production. It has become an indispensable technology for the breeding of new varieties and the promotion of improved varieties, and plays a huge role in the improvement and production of goat varieties. Artificial insemination of sheep is a method in which ram semen is collected mechanically, and after quality inspection and dilution treatment, the semen is injected into the reproductive tract of estrous ewes to replace the natural mating of rams and ewes. The use of artificial insemination has the following advantages: (1) Improve the breeding efficiency of rams, especially in the case of frozen semen, one ram can ma...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12N15/10C12Q1/68
Inventor 凌英会丁建平章孝荣黄桠峰张晓东张子军张运海孙志辉章孝斌
Owner ANHUI AGRICULTURAL UNIVERSITY
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