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  • Title: Rapid detection of male-specific DNA sequence in bovine embryos using fluorescence in situ hybridization.
    Author: Kobayashi J, Sekimoto A, Uchida H, Wada T, Sasaki K, Sasada H, Umezu M, Sato E.
    Journal: Mol Reprod Dev; 1998 Dec; 51(4):390-4. PubMed ID: 9820197.
    Abstract:
    An accurate, reliable, and quick (less than an hour) method for determining the sex of bovine embryos was developed using a fluorescence in situ hybridization (FISH), with a probe designed from a bovine Y chromosome specific DNA (BC1.2). First, to improve a protocol of FISH and evaluate an accuracy of the method, lymphocyte nuclei prepared from three bulls, two cows, and one freemartin were tested. We found that 5 min was enough for hybridization. The washing solution adequate for posthybridization was 0.5x SSC at 72 degrees C for 5 min. The whole procedure for FISH can be accomplished in less than an hour. A male-specific signal was detected, on average, as 97, 0.5, and 83%, respectively, of lymphocytes in males, females, and a freemartin. Using the rapid FISH protocol developed, 28 embryos were divided. According to the presence of the digoxigenin signal, 16 embryos (57.1%) were predicted as male, and 12 embryos (42.9%), predicted as female.
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