title: Comparison of DNA preservation methods for environmental bacterial community samples.
abstract: Field collections of environmental samples, for example corals, for molecular microbial analyses present distinct challenges. The lack of laboratory facilities in remote locations is common, and preservation of microbial community DNA for later study is critical. A particular challenge is keeping samples frozen in transit. Five nucleic acid preservation methods that do not require cold storage were compared for effectiveness over time and ease of use. Mixed microbial communities of known composition were created and preserved by DNAgard(™), RNAlater(®), DMSO-EDTA-salt (DESS), FTA(®) cards, and FTA Elute(®) cards. Automated ribosomal intergenic spacer analysis and clone libraries were used to detect specific changes in the faux communities over weeks and months of storage. A previously known bias in FTA(®) cards that results in lower recovery of pure cultures of Gram-positive bacteria was also detected in mixed community samples. There appears to be a uniform bias across all five preservation methods against microorganisms with high G + C DNA. Overall, the liquid-based preservatives (DNAgard(™), RNAlater(®), and DESS) outperformed the card-based methods. No single liquid method clearly outperformed the others, leaving method choice to be based on experimental design, field facilities, shipping constraints, and allowable cost.
authors.affiliation: U.S. Geological Survey, St. Petersburg, FL, USA.
authors.forename: Michael A
authors.initials: MA
authors.lastname: Gray
authors.forename: Zoe A
authors.initials: ZA
authors.lastname: Pratte
authors.forename: Christina A
authors.initials: CA
authors.lastname: Kellogg
mesh_terms: D001419:Bacteria / Q000235:genetics; D001482:Base Composition; D004269:DNA, Bacterial / Q000737:chemistry; D004783:Environmental Microbiology; D016133:Polymerase Chain Reaction; D011309:Preservation, Biological / Q000379:methods*
chemical_list: D004269:DNA, Bacterial