Method for separating X sperm and Y sperm of pig

A sperm and deionized water technology, applied in the field of separation of pig X sperm and Y sperm, can solve the problems of high cost, insufficient separation purity, influence of vitality and use effect, etc., and achieve the preservation of vitality, high resolution and good vitality Effect

Inactive Publication Date: 2009-12-30
GUANGXI UNIV
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Problems solved by technology

[0004] At present, the technology of separating pig sperm by flow cytometry still has problems such as low sperm separation efficiency, insufficient separation purity, and high cost, and because of the structural problems and separation conditions of pig sperm, the vitality of pig sperm after separation by flow cytometry and the use effect will be affected. So far, there is no mature pig sperm separation method reported in China

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  • Method for separating X sperm and Y sperm of pig
  • Method for separating X sperm and Y sperm of pig
  • Method for separating X sperm and Y sperm of pig

Examples

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Effect test

Embodiment 1

[0039] 1. The freshly collected fine boar semen is quickly transferred to the separation place at 30°C±2°C for temporary storage, and diluted and separated as soon as possible.

[0040] 2. Dilute porcine semen with a volume ratio of 1:1 with semen diluent at an equilibrium temperature (30° C.), and the density of the diluted sperm is 1.15×10 8 / ml.

[0041] 3. Add Hoechst33342 (prepared to a liquid with a concentration of 5 mg / ml) 15 μl / ml (that is, add a fluorescent dye to a concentration of 75 μg / ml) into the diluted pig semen, mix well and place in a water bath at 34°C Stain in a medium water bath with closed light for 50 min. After the water bath is over, add food coloring (FD&C#40) to 25 μg / ml in the fluorescent staining dilute semen.

[0042] Stain at room temperature of 25°C for 5 minutes and then filter to remove low-quality sperm such as agglomerates and dead sperm, and then separate them immediately.

[0043] The composition and content of the semen diluent are:

...

Embodiment 2

[0057] 1. The freshly collected fine boar semen is quickly transferred to the separation place at 30°C±2°C for temporary storage, and diluted and separated as soon as possible.

[0058] 2. Dilute porcine semen with a volume ratio of 2:1 using semen diluent at an equilibrium temperature (30° C.), and the density of the diluted sperm is 0.95×10 8 / ml.

[0059] 3. Add Hoechst33342 (prepared to a liquid with a concentration of 5 mg / ml) 12.5 μl / ml (that is, add a fluorescent dye to a concentration of 62.5 μg / ml) into the diluted pig semen, mix well and store at 37°C Stain in a water bath with closed light for 30 min. After the water bath, add food coloring (FD&C#40) to 20 μg / ml in the diluted semen for fluorescent staining.

[0060] Stain at room temperature of 25°C for 2 minutes and then filter to remove low-quality sperm such as agglomerates and dead sperm, and then separate them immediately.

[0061] The composition and content of the semen diluent are:

[0062] Ele...

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Abstract

The invention relates to a method for separating X sperm and Y sperm of a pig. The method comprises the main steps of: (1) collecting semen of a boar under the temperature of 30+/-2 DEG C and storing temporally; (2) using the semen diluting liquid to dilute the original semen according to the proper proportion, then adding a fluorescent stain with proper concentration, and carrying out light-tight hatching dyeing in a water bath, then adding food pigment for dyeing and then filtering; (3) using a flow cytometer to separate X sperm and Y sperm under the temperature of 20 DEG C to 25 DEG C; and (4) using a receiving pipe filled with sperm receiving liquid to collect the separated X sperm and Y sperm, and being used for storing or inseminating after centrifugation and concentration. The method for separating X sperm and Y sperm of the pig can improve the resolution of the X sperm and Y sperm and separating accuracy rate with the separating accuracy rate of more than 90 percent, and can effectively maintain high quality of sperms and high efficiency of recovery of separated sperms. By being combined with the propagation technologies of artificial insemination, vitro insemination and the like, the method can effectively improve the efficiency of farming industry.

Description

technical field [0001] The invention relates to a method for separating pig X sperm and Y sperm. Background technique [0002] In the breeding industry, selectively breeding a large number of offspring of livestock and poultry whose sex is predicted can greatly improve the production speed and production efficiency. Using X or Y sperm to inseminate female animals can produce many "predicted sex" offspring, which can significantly change the sex ratio of offspring according to our needs, bringing obvious economic and social benefits. For example, in pig production, generally lean pigs have a low reproductive rate and a high lean meat rate, while local breed pigs (such as Taihu pigs and Luchuan pigs) have a high reproductive rate and a low lean meat rate. The male pigs of lean pigs are matched with local sows, which can not only increase the litter size but also increase the lean meat rate of offspring, which is good for improving the economic benefits of farmers and the livi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/06
Inventor 卢克焕曾有权陆阳清张明
Owner GUANGXI UNIV
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