MULTIPLEX POLYMERASE CHAIN-REACTION FOR SEX DETERMINATION OF SINGLE-MOUSE BLASTOMERES

Using a multiplex nested polymerase chain reaction (PCR) method with s ingle copy genes and a dinucleotide repeat locus for the mouse Y and X chromosomes respectively, it was possible to discriminate between sin gle cells derived from male and female embryos, Using single cells, am plification of Sry and Zfy sequences was not evident in all cases, It could be calculated that, with the PCR method used, 0.04% [95% (confid ence interval 0.00-2.03)] of the male embryos would erroneously be dia gnosed as female if analysis is performed on two cells. The calculated chance for total amplification failure, if two cells are used for ana lysis, 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 appl ication of PCR for preimplantation genetic diagnosis at the single cel l level.