A novel method to optimise the utility of underused moulted plumulaceous feather samples for genetic analysis in bird conservation

Autor: Catherine M. Peters, Bonnie L. Rusk, Howard P. Nelson, Anna P. Muir
Rok vydání: 2019
Předmět:
Zdroj: Conservation Genetics Resources. 12:457-467
ISSN: 1877-7260
1877-7252
DOI: 10.1007/s12686-019-01117-8
Popis: Non-invasive sampling methods are increasingly being used in conservation research as they reduce or eliminate the stress and disturbance resulting from invasive sampling of blood or tissue. Here we present a protocol optimised for obtaining usable genetic material from moulted plumulaceous feather samples. The combination of simple alterations to a ‘user-developed’ method, comprised of increased incubation time and modification of temperature and volume of DNA elution buffer, are outlined to increase DNA yield and significantly increase DNA concentration (W = 81, p d = 0.89). We also demonstrate that the use of a primerless polymerase chain reaction (PCR) technique increases DNA quality and amplification success when used prior to PCR reactions targeting avian mitochondrial DNA (mtDNA). A small amplicon strategy proved effective for mtDNA amplification using PCR, targeting three overlapping 314–359 bp regions of the cytochrome oxidase I barcoding region which, when combined, aligned with target-species reference sequences. We provide evidence that samples collected non-invasively in the field and kept in non-optimal conditions for DNA extraction can be used effectively to sequence a 650 bp region of mtDNA for genetic analysis.
Databáze: OpenAIRE
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