Method for efficient ultralow temperature cryopreservation of turbot sperms
An ultra-low temperature freezing and turbot technology, which is applied in the field of marine biology, can solve the problems of restricting the yield of high-quality fertilized eggs and seedlings, low sperm production, short sperm production time of male turbot, etc., and achieves good repeatability and stability. , The effect of large storage capacity and easy operation
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Embodiment 1
[0020] 1) In April 2013, when the broodstock of turbot was in the reproductive period, the male broodstock was fished out from the breeding pond in the broodstock breeding workshop of Jiaonanzeng Breeding Station, Huangdao Island, and placed on a sponge. The reproductive pores were rinsed three times with distilled water, and wiped with paper towels. Clean, gently squeeze the abdomen from back to front to obtain about 30ml of fresh semen from more than ten male fish, store it in a clean centrifuge tube, keep away from light, and the viability of the microscope is higher than 90%, put it in the ice box and bring it back to the experiment room for cryopreservation;
[0021] 2) Antifreeze preparation, which is composed of diluent, antifreeze, additives and distilled water. After preparation, place it in a refrigerator at 0°C for pre-cooling and wait for use; among them, the diluent is: NaCl: 8g / L, KCl: 0.4g / L L, MgSO 4 ·7H 2 O: 0.1g / L, MgCl 2 ·6H 2 O: 0.1g / L, Na 2 HPO 4 2H ...
Embodiment 2
[0030]1) From April to May in 2012, when turbot was in the breeding season, the male fish was taken out of the breeding pond with the broodstock breeding workshop at Jiaonanzeng Breeding Station, Huangdao, and placed on a sponge. The genital pores were rinsed three times with distilled water, and wiped with paper towels. Clean, gently squeeze the abdomen from back to front to get fresh semen. A total of more than 50 tails of turbot were collected, with a total of 70ml of fresh essence, and the viability was 75-90% as detected by the microscope, and they were placed in the ice box and returned to the laboratory for cryopreservation;
[0031] 2) Antifreeze preparation, which is composed of diluent, antifreeze, additives and distilled water. After preparation, place it in a refrigerator at 0°C for pre-cooling and wait for use; among them, the diluent is: NaCl: 8g / L, KCl: 0.4g / L L, MgSO 4 ·7H 2 O: 0.1g / L, MgCl 2 ·6H 2 O: 0.1g / L, Na 2 HPO 4 2H 2 O:0.06g / L,KH 2 PO 4 :0.06g / ...
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