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- Title
Influence of killing method on Lepidoptera DNA barcode recovery.
- Authors
Willows‐Munro, Sandi; Schoeman, M. Corrie
- Abstract
The global DNA barcoding initiative has revolutionized the field of biodiversity research. Such large-scale sequencing projects require the collection of large numbers of specimens, which need to be killed and preserved in a way that is both DNA-friendly and which will keep voucher specimens in good condition for later study. Factors such as time since collection, correct storage (exposure to free water and heat) and DNA extraction protocol are known to play a role in the success of downstream molecular applications. Limited data are available on the most efficient, DNA-friendly protocol for killing. In this study, we evaluate the quality of DNA barcode (cytochrome oxidase I) sequences amplified from DNA extracted from specimens collected using three different killing methods (ethyl acetate, cyanide and freezing). Previous studies have suggested that chemicals, such as ethyl acetate and formaldehyde, degraded DNA and as such may not be appropriate for the collection of insects for DNA-based research. All Lepidoptera collected produced DNA barcodes of good quality, and our study found no clear difference in nucleotide signal strength, probability of incorrect base calling and phylogenetic utility among the three different treatment groups. Our findings suggest that ethyl acetate, cyanide and freezing can all be used to collect specimens for DNA analysis.
- Subjects
LEPIDOPTERA; DNA data banks; BIODIVERSITY; NUCLEOTIDE sequencing; BIOLOGICAL specimens; NUCLEIC acid isolation methods; FORMALDEHYDE
- Publication
Molecular Ecology Resources, 2015, Vol 15, Issue 3, p613
- ISSN
1755-098X
- Publication type
Article
- DOI
10.1111/1755-0998.12331