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DNA extraction techniques for DNA barcoding of minute gall-inhabiting wasps.

Molecular ecology resources (2011-09-29)
Gudrun Dittrich-Schröder, Michael J Wingfield, Hildegard Klein, Bernard Slippers
RÉSUMÉ

DNA extraction from minute hymenopterans and their larvae is difficult and challenging because of their small size indicating a low amount of starting material. Hence, 11 DNA extraction methods were compared to determine their efficacy in isolating DNA. Success of each method was scored on a 2% agarose gel after PCR of the cox 1 mitochondrial locus. A silica-membrane-based approach was the most successful, followed by a method using a combination of incubation buffers and a method using magnetic beads. The method using buffers was the most cost- and time effective. Using this method, larvae from Eucalyptus seed capsule galls could be assigned a role (parasitoid, gall former or inquiline) in the gall-inhabiting complex.

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GenElute Mammalian Genomic DNA Miniprep Kits, sufficient for 350 purifications