Non-invasive sampling: reliable methods for DNA extraction and PCR amplification from a single scale of small Cyprinidae fish

ABSTRACT

The present study was aimed to DNA extraction and PCR amplification of mitochondrial DNA of a varied size range using the non-invasive samples of fish. As many obstacles arise when obtaining an adequate amount of DNA from samples of endangered species, using non-invasive methods of sample collection and DNA extraction should be developed. To this end, we tested and standardised the reliable methods based on DNA extraction from a single scale of small Cyprinidae fish Pethia conchonius. In total, 13 specimens were used for DNA extraction, including 7 single scales and multiple scales. The DNA fragments of different length representing four mitochondrial fragments of different genes were amplified with a variable success after DNA extracted from a single scale, multiple scales or fish clips was used in PCR reactions. All examined DNA samples extracted from a single scale provided 100% amplification success of up to 381 base pairs fragments. Methods described here have a great applicability in DNA extraction from the small size fish and could be useful in conservation genetics as well as in wildlife forensics.

KEYWORDS:

• DNA extraction
• non-invasive samples
• single scale
• Cyprinidae
• wildlife forensics

Acknowledgments

Authors also extent thanks to Director, Dean and Research Coordinator, Wildlife Institute of India, for encouraging this work. The assistance rendered from technical staff from both the laboratories of the University and the Institutes are greatly acknowledged.

Disclosure statement

No potential conflict of interest was reported by the authors.