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Molecular Human Reproduction Vol. 1, NUMBER 1 pp. 45-50, 1995
© European Society of Human Reproduction and Embryology 1995

research-article

Multiplex polymerase chain reaction for sex determination of single mouse blastomeres

J.C.F.M. Dreesen1, J.C.M. Dumoulin1, J.L.H. Evers1, J.P.M. Geraedts2 and M.H.E.C. Pieters1,3

1Department of Obstetrics and Gynaecology, Academic Hospital Maastricht Maastricht, The Netherlands 2Department of Molecular Cell Biology and Genetics, University of Limburg Maastricht, The Netherlands

To whom correspondence should be addressed at: 3To whom correspondence should be addressed at: Academic Hospital Maastricht, Department of Obstetrics and Gynaecology, IVF-laboratory, PO Box 5800, 6202 AZ Maastricht, The Netherlands

Using a multiplex nested polymerase chain reaction (PCR) method with single copy genes and a dinucleotide repeat locus for the mouse Y and X chromosomes respectively, it was possible to discriminate between single cells derived from male and female embryos. Using single cells, amplification of Sry and Zfy sequences was not evident in all cases. ft could be calculated that, with the PCR method used, 0.04% [95% (confidence interval 0.00–2.03)] of the male embryos would erroneously be diagnosed as female if analysis is performed on two cells. The calculated chance for total amplification failure, if two cells are used for analysis, would be 1.4% [95% (confidence interval 0.04–8.04)]. The mouse embryo model proved to be a helpful tool to develop skills in the application of PCR for preimplantation genetic diagnosis at the single cell level.

mouse model/multiplex PCR/preimplantation genetic diagnosis/single cell diagnosis


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