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Molecule diagnosis method for detecting sheep high fecundity gene BMPR1B

A technology of molecular diagnosis and fecundity, applied in biochemical equipment and methods, measurement/testing of microorganisms, DNA/RNA fragments, etc., can solve the problems of increasing detection costs and operating steps, and achieve shortened detection time, reliable results, cost saving effect

Inactive Publication Date: 2013-10-02
赵兴波
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Problems solved by technology

These two methods require the use of special materials such as nucleic acid restriction endonucleases or gene chips, which increase the detection cost and operation steps

Method used

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  • Molecule diagnosis method for detecting sheep high fecundity gene BMPR1B

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specific Embodiment approach

[0006] detailed description The following embodiments will further describe the present invention in detail:

[0007] 1. The total DNA comes from sheep blood, and its extraction method is as follows:

[0008] (1) Add 200 μL of blood sample to a 1.5mL centrifuge tube (EP tube). Add 500ul SNET (1% SDS, 400 mM Nacl, 5 mM EDTA, 20 mM Tris-cl pH8.0), and mix well.

[0009] (2) Add proteinase K to the above solution to a final concentration of 100ng / mL, and mix well.

[0010] (3) Incubate at 55°C for 4 hours.

[0011] (4) Add an equal volume of Tris-saturated phenol, slowly invert the centrifuge tube several times, and centrifuge at 12000g for 10min.

[0012] (5) Take the aqueous phase into a new EP tube, add an equal volume of chloroform / isoamyl alcohol (24:1), shake well for 10 minutes, and centrifuge at 12000g for 10 minutes.

[0013] (6) Take the aqueous phase into a new EP tube, add 1 / 10 volume of 3M NaAc and 2.5 volumes of absolute ethanol, shake well, let stand at -20°...

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Abstract

The present invention discloses a genotype analysis method for sheep high fecundity gene BMPR1B mutation site A746G. According to the present invention, an allele specific PCR method is adopted to design two groups of primers (the first group of the primers comprises common forward primer and wild type reverse primer, and the second group of the primers comprises common forward primer and mutant reverse primer), a genotype of detected sheep is ++ if a 131 bp DNA fragment is only amplified by using the first group of the primers, a genotype of detected sheep is BB if a 131 bp DNA fragment is only amplified by using the second group of the primers, and a genotype of detected sheep is B+ if a 131 bp DNA fragment is amplified by using the first group of the primers and the second group of the primers. With the present invention, uses of restriction endonucleases and DNA chips in the traditional detection method are avoided, the PCR product is directly subjected to agarose gel electrophoresis analysis to obtain a genotype analysis result, and the genotype analysis method is an accurate, simple and rapid sheep high fecundity gene BMPR1B molecule diagnosis method.

Description

technical field [0001] The invention relates to a genotype analysis method of the BMPR1B mutation site A746G of the sheep high fecundity gene. The present invention adopts the method of allele-specific PCR (allele specific PCR, AS-PCR), which can accurately, quickly and easily detect the mutation site A746G of BMPR1B gene, including designing specific primers and genotype analysis according to the mutation site method. Background technique [0002] The fecundity of sheep directly affects the economic benefits of sheep production. Sheep are single-born mammals, but some sheep breeds have stable multiple birth performance, such as Chinese Hu sheep, small-tailed Han sheep, and foreign Booroola Merino sheep (Booroola Merino), Finland Landrace sheep ( Finnish Landrace), Romanov sheep and other breeds. With the development of molecular biology techniques, some efficient molecular markers have been applied to the study of sheep multifetality, and many effective and rapid detecti...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 赵兴波陈晓勇敦伟涛
Owner 赵兴波
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