An efficient protocol for genomic DNA extraction from the endangered species Rhinopithecus brelichi

Abstract

The Guizhou snub-nosed monkey (Rhinopithecus brelichi) is an endangered primate with approximately 750 individuals living in the Mount Fanjing area of northwestern Guizhou province. It is required to understand its evolutionary history and genetic diversity in order to take better measures for protecting this endangered species. As a widely accepted DNA source, faeces of snub-nosed monkeys were collected and used for DNA extraction. We modified the traditional method by washing the intestinal exfoliated cells with acetone and treated the cell lysate with high temperature (just below boiling point at 94 °C). We obtained a high-quality DNA, which was used for polymerase chain reactions (PCRs). Taking the mitochondrial genes (including ND6, tRNA-Glu and Cyto B region) of Sichuan snub-nosed monkey as reference, we performed a PCR and efficiently obtained DNA fragments with expected size. Further sequencing results showed that the amplified mitochondrial gene fragments had a high identity (percentage) with the published gene sequence of Sichuan snub-nosed monkey. These results demonstrated that this modified protocol is reliable and can be used in the genetic studies of monkeys and related species.