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Preservation of corals in salt-saturated DMSO buffer is superior to ethanol for PCR experiments

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Abstract

Specimen collection is time consuming and expensive, yet few laboratories test preservation methods before setting out on field expeditions. The most common preservation buffer used for coral specimens is >70% EtOH. However, alternatives exist that are less flammable, easier to ship, and are widely used in other taxa. Here, we compare the effects of salt-saturated DMSO (SSD) and EtOH preservation buffers on post-extraction DNA quantity and quality. We found that soft tissue integrity was better maintained and higher quantities of DNA were extracted from EtOH-preserved specimens; however, by all other measures, SSD was a superior preservative to EtOH. Extractions of SSD-preserved specimens resulted in higher molecular weight DNA, higher PCR success, and more efficient amplification than specimens preserved in EtOH. Our results show that SSD is generally a superior preservative to EtOH for specimens destined for PCR studies, but species-specific differences indicate that preservation comparisons should be undertaken before collection and storage of samples.

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Acknowledgments

We thank Steve Karl, Sam Kahng, Brian Bowen, members of the ToBo lab, the staff of the Hawai‘i Undersea Research Laboratory (especially Chris Kelley), the staff of NOAA’s Pacific Islands Fisheries Science Center Coral Reef Ecosystem Division (especially Rusty Brainard, Molly Timmers, Megan Moews and Russel Moffit), and participants of the Census of Marine Life ARMS workshop (especially Chris Meyer, Jon Geller, Laetitia Plaisance, Scott Godwin, Gustav Paulay, Julian Caley and Nancy Knowlton) for their intellectual input, feedback, and encouragement to perform this study. We thank the NSF-EPSCoR Evolutionary Genetics facility at HIMB for their assistance in sample processing. This research was funded in part by NSF grant OCE#06-23678, the Hawai‘i Invasive Species Council #53780/53781, the Hawai‘i Coral Reef Initiative, NMSP MOA grant No. 2005-008/66882, and by a grant/cooperative agreement from the National Oceanic and Atmospheric Administration, Project R/HE-1, which is sponsored by the University of Hawai‘i Sea Grant College Program, SOEST, under Institutional Grant No. NA09OAR4170060 from NOAA Office of Sea Grant, Department of Commerce. The views expressed herein are those of the author(s) and do not necessarily reflect the views of NOAA or any of its subagencies. UNIHI-SEAGRANT-JC-09-30. This is contribution #1394 from the Hawai‘i Institute of Marine Biology and #7948 from the School of Ocean and Earth Science and Technology.

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Authors and Affiliations

  1. Hawai‘i Institute of Marine Biology, P.O. Box 1346, Kāne‘ohe, Hawai‘i, 96744, USA

    M. R. Gaither, Z. Szabó, C. E. Bird & R. J. Toonen

  2. Genome and Biomedical Sciences Facility, University of California, Davis, 451 Health Sciences Drive, Davis, CA, 95616, USA

    M. W. Crepeau

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  1. M. R. Gaither
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  2. Z. Szabó
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  3. M. W. Crepeau
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  4. C. E. Bird
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  5. R. J. Toonen
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Corresponding author

Correspondence to M. R. Gaither.

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Communicated by Biology Editor Dr. Ruth Gates

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Gaither, M.R., Szabó, Z., Crepeau, M.W. et al. Preservation of corals in salt-saturated DMSO buffer is superior to ethanol for PCR experiments. Coral Reefs 30, 329–333 (2011). https://doi.org/10.1007/s00338-010-0687-1

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  • DOI: https://doi.org/10.1007/s00338-010-0687-1

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