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Method for detecting sheep FecB gene polymorphism by using Taqman MGB probes

A sheep and probe technology, used in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc. Easy to interpret, easy to interpret, and highly specific effects

Inactive Publication Date: 2015-12-23
INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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Problems solved by technology

Most of the traditional FecB gene detection methods use PCR-RFLP (polymerasechainreaction-restrictionfragmentlengthpolymorphism) and PCR-SSCP (polymerasechainreaction-singlestrandconformationpolymorphism) detection methods. These methods have low throughput and numerous procedures, and it is difficult to achieve high-throughput automated determination

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  • Method for detecting sheep FecB gene polymorphism by using Taqman MGB probes
  • Method for detecting sheep FecB gene polymorphism by using Taqman MGB probes
  • Method for detecting sheep FecB gene polymorphism by using Taqman MGB probes

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Embodiment 1

[0031] Embodiment 1 Utilizes the method for TaqmanMGB probe detection sheep FecB gene polymorphism

[0032] 1. Experimental materials

[0033] A total of 5611 ewes and rams produced by crossbreeding Bamei mutton sheep and small-tailed Han sheep as female parents and other breeds as male parents were selected as the test objects.

[0034] 2. Genomic DNA extraction

[0035] 1 ml of blood was collected from the jugular vein of the sheep and treated with EDTA anticoagulant. First, the red blood cell lysate is lysed to remove DNA-free red blood cells. The nucleus lysate lyses the cells to release genomic DNA. Then the protein precipitation solution selectively precipitates to remove the protein. Finally, the purified genomic DNA is precipitated by isopropanol and redissolved in the DNA lysate.

[0036] 3. Genotyping by TaqmanMGB probe method

[0037] Primers and probes were designed for the 29382188bp site on sheep chromosome 6 (NC_019463.1, based on the sheep genome sequence in...

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Abstract

The invention provides a method for detecting sheep FecB gene polymorphism by using Taqman MGB probes, by which the high-throughput detection of A and G loci can be implemented, and results can be easily interpreted. Compared with methods for detecting FecB genes through traditional PCR-RFLP, and the like, 384 samples can be detected each time, and the following analysis of PCR products is not required to be performed, so that the detection period is greatly shortened and the detection efficiency is improved while the detection cost is saved. Primers and probes used in the invention are strong in specificity, and are not interfered by other genes, so that a risk that false positive is caused by PCR product contaminations is reduced, and the interpretation of results is easier. The high-throughput detection of SNP loci of FecB genes can be implemented, therefore, the method has a potential application value for the large-scale molecular breeding of sheep.

Description

technical field [0001] The invention relates to molecular marker detection technology, in particular to a method for detecting sheep FecB gene polymorphism by using TaqmanMGB probe. Background technique [0002] Livestock litter size is a quantitative trait with enormous economic value. The selection of lamb size is not only restricted by sex and age, but also the lamb size of sheep is a quantitative trait with low heritability, so it is difficult to improve the litter size trait with conventional breeding techniques. Marker-assisted selection can greatly improve the selection efficiency of such low-heritability traits by affecting the selection time, intensity and accuracy of selection, and finding molecular genetic markers linked to these quantitative trait loci is the key to realizing marker-assisted selection. Selected prerequisites. [0003] In the 1950s, the Seears brothers bred a group of Australian Merino (Booroola Merino, BM) that could give birth to multiple birt...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
CPCC12Q1/6851C12Q1/6888C12Q2531/113C12Q2561/101C12Q2561/113
Inventor 储明星刘秋月狄冉胡文萍王翔宇王贵潘章源赵万民李虎山
Owner INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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