Application of an inexpensive and high-throughput genomic DNA extraction method for the molecular ecology of zooplanktonic diapausing eggs

Abstract

We describe the application of a simple, low-cost, and effective method of DNA extraction (hot sodium hydroxide and Tris, HotSHOT) to the diapausing propagules of continental aquatic invertebrates for its use in PCR amplification. We illustrate the use of the technique in cladocerans, rotifers, anostracans, notostracans, and copepod diapausing eggs. We compare the performance of the HotSHOT technique to the currently most widely used method for DNA extraction of zooplankton eggs and individuals, the chelating resin (or Chelex) tech- nique. The HotSHOT technique overcomes several of the problems posed by Chelex and permits easy opti- mization for its use with 96-well plates for high-throughput DNA extraction and subsequent genetic character- ization. We foresee a wide use of this technique in the future from DNA barcoding of diapausing stages to the genetic characterization of the diapausing egg banks of continental aquatic invertebrates.