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Influence of killing method on L epidoptera DNA barcode recovery
Author(s) -
WillowsMunro Sandi,
Schoeman M. Corrie
Publication year - 2015
Publication title -
molecular ecology resources
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.96
H-Index - 136
eISSN - 1755-0998
pISSN - 1755-098X
DOI - 10.1111/1755-0998.12331
Subject(s) - biology , dna , dna extraction , dna sequencing , barcode , computational biology , genetics , polymerase chain reaction , gene , computer science , operating system
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.