Sexing of in vitro fertilized bovine embryos by multiplex PCR

Authors

  • Marcelo Rezende Luz Universidade Estadual Paulista, Faculdade de Ciências Agrárias e Veterinárias, Setor de Reprodução Animal e Obstetrícia, Limoeiro, SP
  • Yeda Fumie Watanabe Universidade de São Paulo, Faculdade de Medicina de Ribeirão Preto, Ribeirão Preto, SP
  • Jesus Aparecido Ferro Universidade Estadual Paulista, Faculdade de Ciências Agrárias e Veterinárias, Setor de Reprodução Animal e Obstetrícia, Limoeiro, SP
  • Maria Inês T. Ferro Universidade Estadual Paulista, Faculdade de Ciências Agrárias e Veterinárias, Setor de Reprodução Animal e Obstetrícia, Limoeiro, SP
  • Sônia Marli Singaretti de Mauro Universidade Estadual Paulista, Faculdade de Ciências Agrárias e Veterinárias, Setor de Reprodução Animal e Obstetrícia, Limoeiro, SP
  • Vera Fernanda Martins Hossepian de Lima Universidade Estadual Paulista, Faculdade de Ciências Agrárias e Veterinárias, Setor de Reprodução Animal e Obstetrícia, Limoeiro, SP
  • Paulo Henrique Franceschini Universidade Estadual Paulista, Faculdade de Ciências Agrárias e Veterinárias, Setor de Reprodução Animal e Obstetrícia, Limoeiro, SP

DOI:

https://doi.org/10.1590/S1413-95962000000600006

Keywords:

Sexing, Embryo, Bovine, PCR

Abstract

In the present study the polymerase chain reaction (PCR) was used for sexing ninety-two in vitro fertilized bovine embryos. The embryos were obtained after in vitro fertilization of oocytes from slaughterhouses. The oocytes were matured, fertilized, and cultured until the blastocyst stage. The embryos were washed in PBS solution, and transferred to polypropylene tubes with containing ultrapure water and immediately frozen at -196ºC. The embryos were thawed on ice and treated with proteinase K. For the PCR reaction, aliquots of 34 µl from each tube were mixed to the primers BC1.2 and microsatellite sequence 1715, dNTPs, MgCl2, 10X PCR buffer, Taq DNA polymerase and water in a final volume of 50 µl. The samples were amplified and the PCR products separated by electrophoresis in a 8% polyacrylamide gel. The gels were stained in ethidium bromide solution and vizualized under UV-light. The amplification rate was 93.47%, with 41 (47.67%) male embryos and 45 (52.32%) female embryos. The use of 8% polyacrylamide gel was efficient for separating DNA fragments of very similar size.

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Published

2000-12-01

How to Cite

Luz, M. R., Watanabe, Y. F., Ferro, J. A., Ferro, M. I. T., Mauro, S. M. S. de, Hossepian de Lima, V. F. M., & Franceschini, P. H. (2000). Sexing of in vitro fertilized bovine embryos by multiplex PCR. Brazilian Journal of Veterinary Research and Animal Science, 37(6), 453-456. https://doi.org/10.1590/S1413-95962000000600006

Issue

Section

VETERINARY MEDICINE

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