Journal
MOLECULAR ECOLOGY RESOURCES
Volume 8, Issue 1, Pages 56-61Publisher
WILEY
DOI: 10.1111/j.1471-8286.2007.01879.x
Keywords
cytochrome c oxidase I; DNA barcoding; DNA extraction; Simuliidae; sonication
Funding
- Natural Environment Research Council [CEH010021] Funding Source: researchfish
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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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