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
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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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