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

Montero-Pau, Javier; Gómez, Africa; Muñoz, Joaquín

Authors

Javier Montero-Pau

Joaquín Muñoz



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) technique. The HotSHOT technique overcomes several of the problems posed by Chelex and permits easy optimization for its use with 96- well plates for high- throughput DNA extraction and subsequent genetic characterization. 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.

Journal Article Type Article
Publication Date 2008-06
Journal LIMNOLOGY AND OCEANOGRAPHY-METHODS
Electronic ISSN 1541-5856
Publisher Wiley
Peer Reviewed Peer Reviewed
Volume 6
Issue 6
Pages 218-222
APA6 Citation Montero-Pau, J., Gómez, A., & Muñoz, J. (2008). Application of an inexpensive and high-throughput genomic DNA extraction method for the molecular ecology of zooplanktonic diapausing eggs. Limnology and oceanography, methods ASLO, 6(6), 218-222. doi:10.4319/lom.2008.6.218
DOI https://doi.org/10.4319/lom.2008.6.218
Keywords DNA extraction; 96 well plates;Genomics
Publisher URL http://www.aslo.org/lomethods/free/2008/0218.pdf