an evaluation of the pcr-rflp technique to aid molecular-based monitoring of felids and canids in indiapcr-rflp技术援助的一个评价分子监测猫科的动物在印度和犬科动物.pdfVIP

Mukherjee et al. BMC Research Notes 2010, 3:159 /1756-0500/3/159 S HO RT REPORT Open Access Short Report An evaluation of the PCR-RFLP technique to aid molecular-based monitoring of felids and canids in India Shomita Mukherjee*, Ashalakshmi CN, Chandrima Home and Uma Ramakrishnan Abstract Background: The order Carnivora is well represented in India, with 58 of the 250 species found globally, occurring here. However, small carnivores figure very poorly in research and conservation policies in India. This is mainly due to the dearth of tested and standardized techniques that are both cost effective and conducive to small carnivore studies in the field. In this paper we present a non-invasive genetic technique standardized for the study of Indian felids and canids with the use of PCR amplification and restriction enzyme digestion of scat collected in the field. Findings: Using existing sequences of felids and canids from GenBank, we designed primers from the 16S rRNA region of the mitochondrial genome and tested these on ten species of felids and five canids. We selected restriction enzymes that would cut the selected region differentially for various species within each family. We produced a restriction digestion profile for the potential differentiation of species based on fragment patterns. To test our technique, we used felid PCR primers on scats collected from various habitats in India, representing varied environmental conditions. Amplification success with field collected scats was 52%, while 86% of the products used for restriction digestion could be accurately assigned to species. We verified this through sequencing. A comparison of costs across the various techniques currently used for scat assignment showed that this technique was the most practical and cost effective. Conclusions: The species-spec