Rapid prenatal sexing of bovine fetus using DNA of fetal fluid cells without prior cell culture
1996
Kadokawa, H. (Hokkaido National Agricultural Experiment Station, Sapporo (Japan)) | Minezawa, M. | Takahashi, H. | Yamada, Y. | Kariya, T.
The objective of this study was to develop rapid prenatal sexing in bovine using DNA of fetal fluid cells and the Polymerase Chain Reaction (PCR) without prior cell culture. In order to archive this objective, the primer pairs were synthesized from the sequence of btDYZ (bovine male specific sequence) and of bActin (bovine specific sequence). PCR was carried out in 100 micro-liter of reaction mixture containing the sample DNA (0.5-4 ng) prepared from 0.5 to 2 ml of fetal fluid. Efficiency of the method was confirmed using both amniotic and allantoic fluids of 23 uteri obtained at a slaughterhouse (13 male and 10 female fetuses of 5 to 36 weeks of age) in preliminary study, i.e., all male fetal fluids were male-specific DNA positive, all female fetal fluids were male-specific DNA negative and all fetal fluids were bovine-specific DNA positive. The fetal fluids were obtained transvaginally from 15 live pregnant cows (with 9 male and 6 female fetuses of 8 to 40 weeks of age) for prenatal sexing and the reliability and feasibility of our procedure were confirmed. Since fetal cells were not necessary required to be alive, allantoic fetal cells could be used in out method. Based on the results from the present study, our procedure could be used as an alternative for the karyotyping of amniotic cells for fetal sexing
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