Application of 2-o-mercapto-acetic indican ascorbic acid in preparation of semen for controlling gender
A technology of ascorbic acid and glucoside, applied in the application field of 2-O-glucoside ascorbic acid in the preparation of sex control semen, to avoid damage and improve sperm motility
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[0018] Example 1
[0019] 1. Take a certain amount of pig semen and dilute it to 200×10 with Hepes buffer 6 Sperm / mL, then add the final concentration of 0.11mM H33342 and the final concentration of 200μM AA-2G (0.068mg / ml), stain for 45min in a 34℃ water bath;
[0020] 2. Further dilute to 100×10 with Hepes buffer containing 4% (g / ml) purified egg yolk and 5% food red (g / ml) 6 Sperm / mL;
[0021] 3. Then use the SX-MoFlo flow cytometer to separate and recover sperm, and detect the collected single-sex sperm to prepare pig sex-controlled semen.
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[0022] Example 2
[0023] 1. Take a certain amount of pig semen and dilute it to 200×10 with Hepes buffer 6 Sperm / mL, then add the final concentration of 0.11mM H33342 and the final concentration of 250μM AA-2G (0.085mg / ml), stain for 45min in a 34℃ water bath;
[0024] 2. Further dilute to 100×10 with Hepes buffer containing 4% (g / ml) purified egg yolk and 5% food red (g / ml) 6 Sperm / mL;
[0025] 3. Then use the SX-MoFlo flow cytometer to separate and recover sperm, and detect the collected single-sex sperm to prepare pig sex-controlled semen.
[0026] Fluorescence microscope was used to detect the semen of pig sex control. The results showed that the acrosome integrity and plasma membrane integrity of sperm were good.
Example Embodiment
[0027] Example 3
[0028] The method is the same as in Example 1, except that the final concentration of AA-2G is adjusted to 150 μM (0.051 mg / ml) to prepare pig sex control semen.
[0029] Fluorescence microscope was used to detect the semen of pig sex control. The results showed that the acrosome integrity and plasma membrane integrity of sperm were good.
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