VERIFICATION OF HORSE MATERNAL LINEAGE BASED ON DERIVED MITOCHONDRIAL-DNA SEQUENCE

Extensive pedigree records in stud books characterize many horse breed s and are a resource for scientific studies, for example testing the c onservation of mitochondrial DNA (mtDNA) nucleotide sequence through e xtended generations. This study showed that even without access to bio logical material from the horses in question, analysis of mtDNA sequen ce in tail-female relatives can be used to derive haplotypes and test alternative dam possibilities. DNA sequence of the D-loop hypervariabl e region was obtained from fragments generated by polymerase chain rea ction (PCR) amplification of mtDNA from peripheral blood samples. PCR primers for the hyperviable region of the D-loop between tRNA(Pro) and the large central conserved sequence block were designed based on pub lished horse sequence. Horses tracing to the same tail-female lines ha d identical DNA sequences, despite separation from common maternal anc estors by more than 80 years and by pedigree loops of up to 17 generat ions.