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

Peters, Catherine
; 
Nelson, Howard
; 
Rusk, Bonnie
; 
Muir, Anna
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Affiliation
University of Chester
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Publication Date
2019-10-24
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Biological Sciences
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Abstract
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 <0.01, Cohens’s 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 314-359bp 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 650bp region of mtDNA for genetic analysis.
Citation
Peters, C., Nelson, H. P., Rusk, B., Muir A. P. (2019) A novel method to optimise the utility of underused moulted plumulaceous feather samples for genetic analysis in bird conservation. Conservation Genetic Resources, 12, 457–467
Publisher
Springer
Journal
Conservation Genetic Resources
Research Unit
URI
http://hdl.handle.net/10034/622766
DOI
10.1007/s12686-019-01117-8
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PubMed Central ID
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Article
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Description
This is a post-peer-review, pre-copyedit version of an article published in Conservation Genetics Resources. The final authenticated version is available online at: http://dx.doi.org/10.1007/s12686-019-01117-8
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ISSN
1877-7252
EISSN
1877-7260
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https://link.springer.com/journal/12686