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RESEARCH ARTICLE
Contents Vol 19(2)

The effects of preservatives and temperatures on arachnid DNA

Cor J. Vink A C , Steven M. Thomas A , Pierre Paquin A , Cheryl Y. Hayashi B and Marshal Hedin A
+ Author Affiliations
- Author Affiliations

A Department of Biology, San Diego State University, San Diego, California 92182-4614, USA.

B Department of Biology, University of California, Riverside, California 92521-0427, USA.

C Corresponding author. Email: cor.vink@arachnology.org

Invertebrate Systematics 19(2) 99-104 https://doi.org/10.1071/IS04039
Submitted: 22 December 2004  Accepted: 12 April 2005   Published: 28 June 2005

Abstract

We tested the effects of different preservatives and temperatures on the yield of spider and scorpion DNA useable for PCR amplification. Our experiment was designed to simulate conditions in the field and laboratory over a six-week time period, testing the preservatives RNAlater®, propylene glycol, and various ethanol concentrations. Three replicates of each preservation treatment were stored at five different temperature treatments; –80°C, –20°C, 2–4°C, 19–24°C, and 40°C. DNA was extracted and quality was assessed by electrophoresis on mini-gels, and by PCR amplification of high copy mitochondrial DNA fragments (cytochrome oxidase subunit I) and low copy nuclear DNA fragments (actin). Results show that RNAlater® and propylene glycol are significantly better than the other preservatives for high quality DNA preservation and that tissue is best stored at –80°C or –20°C. Storage in 95% ethanol is appropriate if specimens are stored at –20°C or –80°C. We believe our results can help guide biologists in choosing preservatives and temperatures for DNA-based research on arachnids, other arthropods and invertebrates in general.

Additional keywords: DNA degradation, DNA preservation, PCR, scorpion, spider.


Acknowledgments

Marie Hudson, Jim Starrett, Ian Ballard and Debra Wytrykush provided valuable assistance with the DNA extractions. Daniel Palmer, Marie Hudson, Jim Starrett, Steve Foldi, Ian Ballard, Erica Dale, Peter Jensen and Stacie Jensen aided with specimen collection. Tom Prentice, Roger Farley, and Rick Vetter provided logistical help. Thanks to Lorenzo Prendini, Jonathan Coddington and Miquel Arnedo for valuable comments during the initial development of this work. We thank Martín Ramírez for his assessment of the morphological condition of spider specimens that had been stored in propylene glycol. Gustavo Hormiga and three anonymous referees provided helpful comments on the manuscript. This research was funded as part of the NSF supported Assembling the Tree of Life: Phylogeny of Spiders, grant number EAR0228699 to Ward Wheeler, Jonathan Coddington, Gustavo Hormiga, Lorenzo Prendini and Petra Sierwald (http://research.amnh.org/atol/files/).


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