An evaluation of the PCR-RFLP technique to aid molecular-based monitoring of felids and canids in India

<p>Abstract</p> <p>Background</p> <p>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 deart...

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Main Authors: Home Chandrima, CN Ashalakshmi, Mukherjee Shomita, Ramakrishnan Uma
Format: Article
Language:English
Published: BMC 2010-06-01
Series:BMC Research Notes
Online Access:http://www.biomedcentral.com/1756-0500/3/159
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author Home Chandrima
CN Ashalakshmi
Mukherjee Shomita
Ramakrishnan Uma
author_facet Home Chandrima
CN Ashalakshmi
Mukherjee Shomita
Ramakrishnan Uma
author_sort Home Chandrima
collection DOAJ
description <p>Abstract</p> <p>Background</p> <p>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.</p> <p>Findings</p> <p>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.</p> <p>Conclusions</p> <p>The species-specific key developed in this paper provides a means for detailed investigations in the future that focus on elusive carnivores in India and this approach provides a model for other studies in areas of Asia where many small carnivores co-occur.</p>
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spelling doaj.art-131fec0f4d92449e9aeeab41d166914f2022-12-22T03:20:31ZengBMCBMC Research Notes1756-05002010-06-013115910.1186/1756-0500-3-159An evaluation of the PCR-RFLP technique to aid molecular-based monitoring of felids and canids in IndiaHome ChandrimaCN AshalakshmiMukherjee ShomitaRamakrishnan Uma<p>Abstract</p> <p>Background</p> <p>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.</p> <p>Findings</p> <p>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.</p> <p>Conclusions</p> <p>The species-specific key developed in this paper provides a means for detailed investigations in the future that focus on elusive carnivores in India and this approach provides a model for other studies in areas of Asia where many small carnivores co-occur.</p>http://www.biomedcentral.com/1756-0500/3/159
spellingShingle Home Chandrima
CN Ashalakshmi
Mukherjee Shomita
Ramakrishnan Uma
An evaluation of the PCR-RFLP technique to aid molecular-based monitoring of felids and canids in India
BMC Research Notes
title An evaluation of the PCR-RFLP technique to aid molecular-based monitoring of felids and canids in India
title_full An evaluation of the PCR-RFLP technique to aid molecular-based monitoring of felids and canids in India
title_fullStr An evaluation of the PCR-RFLP technique to aid molecular-based monitoring of felids and canids in India
title_full_unstemmed An evaluation of the PCR-RFLP technique to aid molecular-based monitoring of felids and canids in India
title_short An evaluation of the PCR-RFLP technique to aid molecular-based monitoring of felids and canids in India
title_sort evaluation of the pcr rflp technique to aid molecular based monitoring of felids and canids in india
url http://www.biomedcentral.com/1756-0500/3/159
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