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

Authors

  • GUDRUN DITTRICH-SCHRÖDER,

    1. Forestry and Agricultural Biotechnology Institute, University of Pretoria, Pretoria 0002, South Africa
    2. Department of Zoology and Entomology, University of Pretoria, Pretoria 0002, South Africa
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  • MICHAEL J. WINGFIELD,

    1. Forestry and Agricultural Biotechnology Institute, University of Pretoria, Pretoria 0002, South Africa
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  • HILDEGARD KLEIN,

    1. ARC-Plant Protection Research Institute, Queenswood, Pretoria 0121, South Africa
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  • BERNARD SLIPPERS

    1. Forestry and Agricultural Biotechnology Institute, University of Pretoria, Pretoria 0002, South Africa
    2. Department of Genetics, University of Pretoria, Pretoria 0002, South Africa
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Prof Bernard Slippers, Fax: +27124203960; E-mail: Bernard.Slippers@fabi.up.ac.za

Abstract

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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