Validating FTA card skin swabs as a source of DNA from wild amphibians

Ward, Ashlee 2016, Validating FTA card skin swabs as a source of DNA from wild amphibians , MSc by research thesis, Salford University.

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Abstract

Amphibians are the most endangered groups of vertebrates, and there is currently an urgent need to implement efficient monitoring schemes based on insights from DNA. However, largely due to their unprotected skin which lacks dead keratinised tissue, current standard techniques for DNA sampling of endangered amphibians are either destructive (sacrifice of entire animals) or can involve distinctly invasive techniques such as tissue sampling based on toe or tail tip clips. The present MSc by Research for the first time quantitatively investigates whether skin swabs with commercially available Whatman FTA cards, a relatively uninvasive technique, can yield DNA samples of sufficient quality and quantity for PCR amplification. In total, swabs from 22 individual great crested newts (Triturus cristatus, the most endangered newt in the UK and an important flagship species for conservation) were analysed, and partly directly compared with toe clip samples. For amplification tests, 7 nuclear microsatellite loci and one mitochondrial locus (ND4) were used; the success rates of PCRs were quantified by replicate PCRs based on DNA extractions using different protocols. While PCR success rates differed across genetic markers, the study showed that, with sufficient replicates, it is possible to efficiently retrieve amplifiable amphibian DNA based on FTA-card skin swabs. The most successful protocol was based on the merging of eight punches from different areas of the FTA card for DNA extraction using the commercially available Qiagen kit, leading to PCR amplification rates which were largely indiscernible from PCRs based on tissue samples. The established new protocol for DNA collection should aid in reducing the invasiveness of DNA collection in future amphibian studies.

Item Type: Thesis (MSc by research)
Schools: Schools > School of Environment and Life Sciences
Funders: Non funded research
Depositing User: Ashlee Ward
Date Deposited: 08 Dec 2016 13:43
Last Modified: 08 Dec 2016 13:43
URI: http://usir.salford.ac.uk/id/eprint/40085

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