Sheep X and Y sperm sorting liquid, sorting floating system and sorting method

A technology for sorting liquid and sperm, which is applied in the field of Y sperm sorting and mammalian X, can solve the problems that are difficult to apply, and achieve the effect of high separation efficiency, low separation cost and good results

Pending Publication Date: 2021-05-28
NORTHWEST A & F UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the biggest problem at present is that the use of flow cytometry to produce sex-controlled semen has

Method used

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  • Sheep X and Y sperm sorting liquid, sorting floating system and sorting method
  • Sheep X and Y sperm sorting liquid, sorting floating system and sorting method
  • Sheep X and Y sperm sorting liquid, sorting floating system and sorting method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] The formulation of X and Y sperm sorting liquid of this embodiment is: glucose 27g / L, citric acid 0.45g / L, lycopene 0.2g / L, sodium citrate 1.9g / L, EDTA 0.3g / L, bicarbonate Sodium 0.9g / L, potassium chloride 0.7g / L, Tris 5.6g / L, BSA 0.8g / L, ultrapure water 1L.

[0035] The formula of the semen dilution washing liquid is: Tris 200mM, citric acid 80mM, fructose 65mM, ultrapure water 100mL.

Embodiment 2

[0037] The formulation of X and Y sperm sorting liquid of this embodiment is: glucose 31g / L, citric acid 0.8g / L, lycopene 0.6g / L, sodium citrate 2.3g / L, EDTA 0.7g / L, bicarbonate Sodium 1.3g / L, potassium chloride 1.1g / L, Tris 6.0g / L, BSA0.5g / L, ultrapure water 1L.

[0038] The formula of semen dilution and washing liquid is: Tris 300mM, citric acid 90mM, fructose 75mM, ultrapure water 100mL.

Embodiment 3

[0040] In this embodiment, the sorting floating system is used to sort the X and Y sperm of dairy goats, and the formula of the sheep semen diluted washing liquid used is: Tris 250mM, citric acid 85mM, fructose 70mM, ultrapure water 100mL;

[0041] The formula of X and Y sperm sorting liquid used is: glucose 27.5g / L, citric acid 0.45g / L, lycopene 0.4g / L, sodium citrate 2.1g / L, EDTA 0.4g / L, sodium bicarbonate 1.2 g / L, potassium chloride 0.9g / L, Tris 5.8g / L, BSA 0.4g / L, ultrapure water 1L.

[0042] The specific sorting method is as follows:

[0043] (1) Wash the semen 3 times with 3mL semen diluted washing solution (800rpm, 5min / time); then take 3mL semen washing solution, first dilute the washed sperm sample, the concentration is about 1×10 8 Sperm / mL, then centrifuge twice with 3mL diluted washing solution (800rpm, 5min / time), and remove the supernatant;

[0044] (2) Resuspend the sperm sample pellet in 3mL semen sorting solution containing 0.3μM R848, and then incubate the ...

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Abstract

The invention discloses sheep X and Y sperm sorting liquid, a sorting floating system and a sorting method. According to the disclosed scheme, sheep sperms X and Y are sorted by using a specific protein TLR7/8 agonist R848 "floating system". According to the technology for activating and sorting the X and Y sperms through the specific protein TLR7/8 of the X and Y sperms of the milk goat, the separation method is simple, the effect is good and reliable, and the X and Y sperms of the milk goat can be efficiently separated; the X sperms sorted by the method are used for in-vitro fertilization, and the proportion of female embryos reaches 80.52%; the X sperms sorted by the method are used for artificial insemination, and the proportion of female embryos reaches 88.89%.

Description

technical field [0001] The invention relates to mammalian X and Y sperm sorting technology, in particular to sheep X and Y sperm sorting solution, sorting system and sorting method. [0002] technical background [0003] In animal husbandry production, sex control technology is to make female animals produce offspring of a specific sex according to people's wishes through certain interventions. Through gender control, the economic benefits of animal husbandry can be improved; serial reproductive diseases can be avoided through artificial insemination technology, embryo transfer technology, etc., thereby speeding up the breeding and breeding process of livestock and poultry. The gender control of sheep, such as dairy goats, is the process of artificially controlling the breeding of dairy goats. It is mainly to effectively separate X and Y sperm, and use the separated sperm to artificially inseminate ewes, so as to produce dairy goat offspring that meet the required gender . ...

Claims

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Application Information

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IPC IPC(8): C12N5/076
CPCC12N5/061C12N5/0612C12N2509/00
Inventor 胡建宏任发任毅杰李宇席华明温飞贤明赵梦洁朱大伟王立强雷安民李延华
Owner NORTHWEST A & F UNIV
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