Nondestructive DNA extraction from blackflies (Diptera: Simuliidae): retaining voucher specimens for DNA barcoding projects |
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| Authors: | Hunter Stephanie J Goodall Tim I Walsh Kerry A Owen Richard Day John C |
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| Affiliation: | Department of Biochemistry and Molecular Biology, University College London, Gower Street, London WC1E 6BT, UK, NERC Centre for Ecology and Hydrology - Oxford, Mansfield Road, Oxford OX1 3SR, UK, Science Department, Environment Agency, Evenlode House, Howberry Park, Wallingford OX10 8BD, UK. |
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| Abstract: | 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. |
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| Keywords: | cytochrome c oxidase I DNA barcoding DNA extraction Simuliidae sonication |
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