SEX DETERMINATION OF IN-VITRO DEVELOPED BUFFALO (BUBALUS-BUBALIS) EMBRYOS BY DNA AMPLIFICATION

This study was conducted to determine the sex of buffalo embryos produ ced in vitro by amplifying male specific DNA sequences using the polym erase chain reaction (PCR). This method uses three different pairs of bovine Y-chromosome specific primers and a pair of bovine satellite sp ecific primers. Buffalo in vitro fertilized embryos at the 4-cell to b lastocyst stage were collected at days 3, 4, 6, and 8 postinsemination , and the sex of each embryo was determined using all three different Y-chromosome specific primers. The bovine satellite sequence specific primers recognize similar sequences in buffalo and are amplified both in males and in females. Similarly, Y-chromosome specific primers ampl ify the similar Y-chromosome specific sequences in male embryos of buf falo. Upon examining genomic DNA from lymphocytes of adult males and f emales, and embryos, the results demonstrate the feasibility of embryo sexing in buffaloes. Furthermore, sex determination by PCR was found to be a rapid and accurate method. (C) 1993 Wiley-Liss, Inc.