Method for detecting sheep FecB gene polymorphism through Taqman MGB probe

A sheep and probe technology, used in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of difficult high-throughput automated measurement, low throughput, and many procedures, and achieve easy and specific interpretation of results. Strong, easy to read effect

Inactive Publication Date: 2015-10-07
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 (polymerase chain reaction-restriction fragment length polymorphism) and PCR-SSCP (polymerase chain reaction-single strand conformation polymorphism) detection methods. Difficult to achieve high-throughput automated assays

Method used

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  • Method for detecting sheep FecB gene polymorphism through Taqman MGB probe
  • Method for detecting sheep FecB gene polymorphism through Taqman MGB probe
  • Method for detecting sheep FecB gene polymorphism through Taqman MGB probe

Examples

Experimental program
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Effect test

Embodiment 1

[0031] Embodiment 1 Utilizes the method for Taqman MGB 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 the female parent and other breeds as the male parent were selected as the test objects.

[0034] 2. Genomic DNA extraction

[0035] 1ml 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 Taqman MGB probe method

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

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Abstract

The invention provides a method for detecting sheep FecB gene polymorphism through a Taqman MGB probe. High throughput detection on A and G sites can be achieved, and results can be easily judged and read. Compared with a traditional method for detecting FecB genes through PCR-RFLP and the like, 384 samples can be detected each time; follow-up analysis of PCR products is not needed; the detection period is greatly shortened while the detection cost is saved; and the detection efficiency is improved. Primers and the probe used in the method are high in specificity; interferences of other genes can be avoided; false positive risks caused by PCR product pollution are reduced; and the result judgment and reading are easier. High throughput detection on SNP sites of the FecB genes can be carried out, and the method has the potential application value in sheep large-scale molecular breeding.

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 Taqman MGB 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 bir...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
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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