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Method and kit for identifying early embryonic sex of pig by dual temperature multiplex PCR

A kit and embryo technology, applied in the PCR reaction technology system, dual-temperature multiplex PCR to identify the gender of early pig embryos, can solve the problems of unseen pig early embryo sex identification methods, and achieve cost-saving, economical and convenient, and sample-saving The effect of using

Inactive Publication Date: 2010-06-09
JILIN UNIV
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Problems solved by technology

[0003] At present, the sex identification methods of livestock mainly include cytogenetic method, H-Y antigen method, sex-specific DNA probe method, PCR method, LAMP method, etc., which are mainly used for sex identification of cattle, sheep or other economic animals. There are literature reports on the method of sex identification of early pig embryos

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  • Method and kit for identifying early embryonic sex of pig by dual temperature multiplex PCR

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Experimental program
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Effect test

Embodiment 1

[0045] 1. Sample handling:

[0046] A: If the sample to be tested is pig embryo or cell:

[0047] Separate 4 or more cells from the early pig embryos to be tested, put them into a 0.2ml PCR tube, add 10ul of cell lysate, place in a PCR instrument at 56°C for 10min, 94°C for 5min; centrifuge at 12000rpm for 2min, and take the supernatant , stored at -20°C for later use;

[0048] B: If the sample to be tested is pig early fetus or pig tissue of unknown sex:

[0049] Take the 1mm×1mm size tissue block of the sample to be tested, put it into a 0.2ml PCR tube, add 50ul of tissue lysate, place it in a PCR instrument at 56°C for 50min, and at 94°C for 5min; centrifuge at 12000rpm for 2min, take the supernatant and store it at -20°C for later use ;

[0050] 2. Primer design:

[0051] A pair of dual-temperature PCR amplification primers were designed according to the porcine SRY gene sequence (Genbank accession number: U49860.2). In addition, a pair of dual-temperature PCR amplifi...

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Abstract

The invention discloses a method and a kit for identifying the early embryonic sex of a pig by dual temperature multiplex PCR. A dual temperature multiplex PCR reaction system with rapidness, sensitiveness and high accuracy, which is suitable for identifying the early embryonic sex of the pig, is used for identifying the early embryonic sex of the pig. The kit which is developed by the dual temperature multiplex PCR reaction system and used for identifying the early embryonic sex of the pig fully plays the characteristics of high efficiency, economy, simpleness, convenience and high reaction specificity of the dual temperature multiplex PCR and realizes the rapid identification of the early embryo of the pig, thereby realizing the sex control of the pig, improving the economic benefits ofpig breeding industry, saving the breeding cost and providing convenience.

Description

technical field [0001] The invention relates to a PCR reaction technology system suitable for sex identification of early pig embryos, and further discloses a double-temperature multiplex PCR method for sex identification of early pig embryos, and also provides corresponding kits, belonging to livestock sex control Identification of technical fields. Background technique [0002] The sex control of domestic animals refers to the technology of making female breeding animals reproduce the desired sex offspring according to people's wishes through human intervention, which is of great significance in animal husbandry production. [0003] At present, the sex identification methods of domestic animals mainly include cytogenetic method, H-Y antigen method, sex-specific DNA probe method, PCR method, LAMP method, etc., which are mainly used for sex identification of cattle, sheep or other economic animals. There are reports in the literature on methods for sex identification of ear...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 欧阳红生李小平逄大欣高飞闫森赖良学王铁东李莉于浩
Owner JILIN UNIV
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