Abstract
Abstract A procedure for multilocus genetic typing (sex and four genetic diseases) of bovine preimplantative embryos using multiplex PCR is described. One hundred sixteen day 6 to 7 embryos were micromanipulated to isolate 5 to 20 blastomeres transferred into a reaction tube containing 10 μl of distilled water. The subsequent genetic typing involved DNA release by proteinase K‐treatment, simultaneous amplification of bovine DNA sequences at 5 different loci ‐ a male specific DNA‐sequence, thyroglobulin gene (congenital hypothyroidism), argininosuccinate synthetase gene (citrullinaemia), CD‐18 gene (BLAD) and uridine monophosphate synthase gene (DUMPS) ‐ and analysis of each genetic variant by subsequent monolocus PCR followed by electrophoretic characterization of PCR or RFLP fragments. Successful genetic analysis was performed in 90 to 100% of the embryos with respect to sex and genetic disease determination.
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