A reliable non-invasive PCR method for takin (Budorcas taxicolor) sex identification based on amelogenin gene

Abstract

We developed a reliable non-invasive PCR method for sex identification of the takin (Budorcas taxicolor) based on Amelogenin (AMEL) genes. In takin, a 45 bp deletion of Y-linked allele provides a significant distinction between AMELX and AMELY, and amplification products show sex-specific banding patterns (male: 240 and 195 bp; female: 240 bp) after agarose gel electrophoresis. However, SRY gene produces only male-specific amplification, and amplification products have only one band (male: 230 bp). Both feces and muscle samples from known-sex takins were successfully amplified. There was no amplification failure of our specific primers, and the phenotypic and genotypic of 15 tested taxins (4 males and 11 females) were highly consistent. Cross-species detection also proved that our primers could be applied to other Caprinae species. The detection sensitivity of this method was 50 pg of genomic DNA. These results show that using one pair of primers to co-amplify homologous fragments according to the AMEL gene exon 5 is a reliable and rapid method.