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Comparison of DNA preservation methods for environmental bacterial community samples

FEMS Microbiology Ecology

By:
, ,
DOI: 10.1111/1574-6941.12008

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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.

Additional Publication Details

Publication type:
Article
Publication Subtype:
Journal Article
Title:
Comparison of DNA preservation methods for environmental bacterial community samples
Series title:
FEMS Microbiology Ecology
DOI:
10.1111/1574-6941.12008
Volume
83
Issue:
2
Year Published:
2013
Language:
English
Publisher:
Wiley
Publisher location:
Hoboken, NJ
Contributing office(s):
St. Petersburg Coastal and Marine Science Center
Description:
10 p.
Larger Work Type:
Article
Larger Work Subtype:
Journal Article
Larger Work Title:
FEMS Microbiology Ecology
First page:
468
Last page:
477