DNA extraction protocols for molecular studies of Asiatic black bears

Abstract

Abstract Optimization of DNA extraction and amplification techniques increases the reliability of genetic studies used to estimate population size and assess genetic diversity, particularly for non-invasively collected samples. We evaluated the effectiveness of 4 DNA extraction procedures—the Roboscreen and Qiagen kit methods and the Organic and Chelex methods, as applied to invasively and non-invasively collected samples from Asiatic black bears (Ursus thibetanus). We assessed effectiveness based on quantity and purity of the extracts by method, and their ability to amplify genomic DNA via polymerase chain reaction (PCR) amplification of a single simple sequence repeat (SSR) marker. For each method, the greatest average DNA quantity and purity index (86.25%) was from blood, while muscle showed greatest DNA quantity and purity index using the Qiagen DNA extraction method. The Roboscreen kit and Chelex method provided a 57% and 58% PCR success rate, respectively, for hairs, demonstrating the value of such ...