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Nondestructive DNA extraction from blackflies (Diptera: Simuliidae): retaining voucher specimens for DNA barcoding projects

Hunter, Stephanie J.; Goodall, Tim I.; Walsh, Kerry A.; Owen, Richard; Day, John C.. 2008 Nondestructive DNA extraction from blackflies (Diptera: Simuliidae): retaining voucher specimens for DNA barcoding projects. Molecular Ecology Resources, 8. 56-61. 10.1111/j.1471-8286.2007.01879.x

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Abstract/Summary

A nondestructive, chemical-free method is presented for the extraction of DNA from small insects. Blackflies were submerged in sterile, distilled water and sonicated for varying lengths of time to provide DNA which was assessed in terms of quantity, purity and amplification efficiency. A verified DNA barcode was produced from DNA extracted from blackfly larvae, pupae and adult specimens. A 60-second sonication period was found to release the highest quality and quantity of DNA although the amplification efficiency was found to be similar regardless of sonication time. Overall, a 66% amplification efficiency was observed. Examination of post-sonicated material confirmed retention of morphological characters. Sonication was found to be a reliable DNA extraction approach for barcoding, providing sufficient quality template for polymerase chain reaction amplification as well as retaining the voucher specimen for post-barcoding morphological evaluation.

Item Type: Publication - Article
Digital Object Identifier (DOI): 10.1111/j.1471-8286.2007.01879.x
Programmes: CEH Programmes pre-2009 publications > Biodiversity > BD03 The Genetic Basis of Ecological Function > BD03.3 Molecular evolution and bioinformatics
CEH Programmes pre-2009 publications > Biodiversity > BD03 The Genetic Basis of Ecological Function > BD03.1 Molecular ecology
CEH Sections: Hails
ISSN: 1755-098X
Additional Keywords: cytochrome c oxidase I, DNA barcoding, DNA extraction, Simuliidae, sonication
NORA Subject Terms: Zoology
Biology and Microbiology
Date made live: 14 Jan 2008 15:07
URI: http://nora.nerc.ac.uk/id/eprint/1911

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