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Establishing and optimizing the system for sex determination of bovine preimplantation embryos

Published online by Cambridge University Press:  12 February 2007

Chen Cong-Ying
Affiliation:
Key Laboratory for Animal Biotechnology of Jiangxi Province and the Ministry of Agriculture, Jiangxi Agricultural University, Nanchang, 330045, China
Huang Lu-Sheng*
Affiliation:
Key Laboratory for Animal Biotechnology of Jiangxi Province and the Ministry of Agriculture, Jiangxi Agricultural University, Nanchang, 330045, China
Chen Jing-Bo
Affiliation:
Institute of Animal Science, Xinjiang Academy of Animal Science, Urumqi, 830000, China
Ding Neng-Shui
Affiliation:
Key Laboratory for Animal Biotechnology of Jiangxi Province and the Ministry of Agriculture, Jiangxi Agricultural University, Nanchang, 330045, China
Zhou Li-Lua
Affiliation:
Key Laboratory for Animal Biotechnology of Jiangxi Province and the Ministry of Agriculture, Jiangxi Agricultural University, Nanchang, 330045, China
Xiao Hai-Xia
Affiliation:
Institute of Animal Science, Xinjiang Academy of Animal Science, Urumqi, 830000, China
*
*Corresponding author. E-mail: [email protected]

Abstract

Two pairs of nested primers were derived from the 5′ flanking sequence of the bovine SRY gene to serve as male-specific sexing primers, which were used for PCR-based sex determination of bovine embryos in combination with casein protein gene primers as internal control. At the same time, methods of DNA extraction from bovine embryo cells were established and the sensitivity of nested PCR and normal PCR was studied for developing a technique of sex determination of bovine embryos adapted for practical use. Results showed that the primers used in the study were male-specific and all primers were bovine-specific. Boiling and freeze–thawing were used for DNA extraction from embryo cells. Amplification products were obtained with only 10 cells by nested PCR, so this system was used for the sex determination of bovine embryos. The sex of 10 Holstein cow embryos was identified and the results were confirmed upon birth of the calves.

Type
Research Article
Copyright
Copyright © China Agricultural University and Cambridge University Press 2004

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