A Non-Invasive Approach to The Genetic Analysis of Asiatic Black Bear

Genetic analysis of mammalian free-ranging populations and endangered species utilizing molecular methods,such as protein electrophoresis,DNA RFLP and DNA sequencing,has long been hampered by tissue-sampling problems.We established a non-invasive approach to help to overcome the problems.The genomic DNA of the Asiatic black bear was extracted from single hair shed on the ground using InstaGene Purification Matrix (Bio-rad) with a simple cell lysis step by boiling in the presence of the matrix.Our protocols allow simple,easy and fast DNA preparation by eliminating labor intensive phenol/chloroform extraction.Mitochondrial cytochrome b gene fragment was amplified from hair DNA using PCR technique.Double stranded PCR product was directly sequenced by the dideoxy method using heat denaturation.Our success on determination of mitochondrial cytochrome b gene sequence from hair could be considered as a first step in the population genetic study of the free-ranging Asiatic black bears.