DNA extraction from Eois (Geometridae) moths - Bertin Instruments

    Environment

    DNA extraction from Eois (Geometridae) moths

    Sources: Department of Animal Biodiversity, Univ. of Vienna, Austria.

    Context

    DNA barcoding uses a standardized mitochondrial DNA fragment to distinguish and identify species. Individual specimens of small sized moths have to be extracted. In order to save as much as possible for morphological analyses the amount of material available for DNA extraction is often very small. Insects collected in the field are usually in less than optimal condition for DNA extraction (i.e. DNA may be partially degraded). A suitable method for extraction needs to provide fast, easy and contamination free homogenization of tissue samples. Specimens are usually irreplaceable and therefore highly valuable, homogenization methods need to conform to high standards of reliability, reproducibility and efficient use of even the smallest amount of sample.[1]

    [1] Strutzenberger, P., Brehm, G. and Fiedler, K. ,DNA barcoding-based species delimitationincreases species count of Eois (Geometridae)moths in a well-studied tropical mountain forestby up to 50%. Insect Science, no. doi:10.1111/j.1744-7917.2010.01366.

    Results

    Figure 1 shows an agarose gel loaded with extracted DNA of Eois moths homogenized with Precellys®24. High quality DNA (>10000bp) could be obtained from all extracted specimens. Tissue lysis with Precellys®24 is very efficient and time saving as subsequent proteinase digestion can be drastically shortened. Precellys homogenizers provide fast and cross contamination safe DNA extraction. Tissue homogenization with Precellys®24 provided the maximum possible DNA yield from small sized moths as used in this study.

    1

    Figure 1: Agarose gel electrophoresis of extracted specimens Lane 1: Size marker; lanes 2-13: DNA extracted from legs

     

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