Abstract
This report describes BioDry (patent pending), a method for reliably preserving the biomolecules associated with aquatic microbial biomass samples, without the need of hazardous materials (e.g. liquid nitrogen, preservatives, etc.), freezing, or bulky storage/sampling equipment. Gel electrophoresis analysis of nucleic acid extracts from samples treated in the lab with the BioDry method indicated that molecular integrity was protected in samples stored at room temperature for up to 30 days. Analysis of 16S/18S rRNA genes for presence/absence and relative abundance of microorganisms using both 454-pyrosequencing and TRFLP profiling revealed statistically indistinguishable communities from control samples that were frozen in liquid nitrogen immediately after collection. Seawater and river water biomass samples collected with a portable BioDry “field unit", constructed from off-the-shelf materials and a battery-operated pumping system, also displayed high levels of community rRNA preservation, despite a slight decrease in nucleic acid recovery over the course of storage for 30 days. Functional mRNA and protein pools from the field samples were also effectively conserved with BioDry, as assessed by respective RT-PCR amplification and western blot of ribulose-1-5-bisphosphate carboxylase/oxygenase. Collectively, these results demonstrate that BioDry can adequately preserve a suite of biomolecules from aquatic biomass at ambient temperatures for up to a month, giving it great potential for high resolution sampling in remote locations or on autonomous platforms where space and power are limited.
Highlights
Discerning the dynamic processes and interactions of natural microbial communities is key to understanding global biogeochemical cycles
There was a slight, but repeatable decrease in differential air-flows (DNA) yield after samples were treated with BioDry, but no sign of degradation or shearing of the nucleic acids was apparent
In all molecular integrity/contamination tests, PCR amplicon was detected for the seawater biomass samples, and no PCR amplicon was detected for the “air-blank” filters
Summary
Discerning the dynamic processes and interactions of natural microbial communities is key to understanding global biogeochemical cycles. Efforts over the past two decades have revealed that microbial communities are more diverse, complex, and variable than previously appreciated for nearly every environment that has been investigated [1,2]. There is increasing evidence that single-celled populations can exhibit functional variability over small. BioDry: A New Method for Preserving Microbial Biomass study design, data collection and analysis, decision to publish, or preparation of the manuscript
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