title: Sex determination and milk protein genotyping of preimplantation stage bovine embryos using multiplex PCR.
abstract: A method for determining the sex and milk protein genotypes (RFLPs) of preimplantation stage bovine embryos using multiplex polymerase chain reaction (PCR) is described. Day 6 to 7 embryos were micromanipulated to isolate 5 to 6 cells. These cells were then dried in reaction tubes for transport to the laboratory. Subsequently, two sets of PCRs were performed using Y chromosome, k-casein and beta-lactoglobulin gene specific primers, followed by electrophoretic analysis of the PCR products. The presence or absence of the Y chromosome was ascertained in 90 of 92 embryos. Moreover, the k-casein specific fragment was amplified and detected in all these embryos. The PCR products were digested in order to genotype the k-casein gene. In 70% of the embryos, the beta-lactoglobulin specific fragment was amplified, although together with some unspecific fragments.
authors.affiliation: Institute of Animal Husbandry and Breeding, Deptartment of Animal Breeding (470), University of Hohenheim, 7000 Stuttgart 70, Germany.
authors.forename: P L
authors.initials: PL
authors.lastname: Agrawala
authors.forename: V A
authors.initials: VA
authors.lastname: Wagner
authors.forename: H
authors.initials: H
authors.lastname: Geldermann