Application of an inexpensive and high-throughput genomic DNA extraction method for the molecular ecology of zooplanktonic diapausing eggs
Application of an inexpensive and high-throughput genomic DNA extraction method for the molecular ecology of zooplanktonic diapausing eggs
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DOI：
10.4319/lom.2008.6.218
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发表时间：
2008-06-01
影响因子：
2.7
通讯作者：
Munoz, Joaquin
中科院分区：
文献类型：
--
作者：
Montero-Pau, Javier;Gomez, Africa;Munoz, Joaquin
本文介绍了一种简单、低成本、有效的DNA提取方法(热氢氧化钠和Tris，Hotshot)在大陆水生无脊椎动物滞育繁殖中的应用，以用于PCR扩增。我们举例说明了这项技术在枝角类、轮虫、无尾轮虫、无尾轮虫和桡足类滞育卵中的应用。我们比较了HotShot技术和目前最广泛使用的浮游动物卵和个体DNA提取方法--螯合树脂(或Cherex)技术的性能。Hotshot技术克服了CHILEX提出的几个问题，并使其与96孔板一起用于高通量DNA提取和随后的基因表征而易于优化。从滞育阶段的DNA条形码到大陆水生无脊椎动物滞育卵库的遗传特征，我们预计这项技术将在未来得到广泛的应用。
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.