Fig 1.
Agarose gel of DNA/RNA extracts from the laboratory samples preserved with BioDry.
The frozen controls (C1 and 2) are followed by samples that were preserved at either high or low air-flow rates, for 10 (lanes 10a and b) or 20 (lanes 20a and b) min, and then stored at room temperature in a desiccant chamber for T15 or T30 days (top & bottom images respectively). Lanes E1 and 2 in the bottom image represent the filters whose holders were exposed to ambient air for 2 days.
Table 1.
Biomolecule Quantification (±SD)–Nucleic acid quantity (ng/μl) from laboratory tests in controls vs. BioDried samples (15d or 30d); Field tests of nucleic acids (ng/μl), total protein content (ng/μl), and RuBisCo large subunit (fmol/μg of protein by Western Analysis) from BioDried samples (10d or 30d; na = not available).
Table 2.
OTU and Sorensen’s similarity index measure (CS) for the laboratory verification tests.
The average number of bacterial OTUs (± SD) are indicated. (There is no SD for the pooled DNA-TRFLP or 454 samples; for RNA-TRFLP the n = 2 for controls and n = 8 for preserved; na = not available).
Fig 2.
Comparison of species relative percent contributions in frozen controls with samples preserved using BioDry, as determined by 454-pyrosequencing.
Results shown for the comparison of DNA samples (n = 3 per treatment) from BioDried samples that were stored for T15 and T30 d (black and blue profiles respectively) with frozen/control samples. Values represent only those peaks present in more than one replicate for the treatments being compared; the number of OTUs used for each analysis is indicated at the bottom-right of each graph; see S2 Table for taxonomy and relative contributions.
Fig 3.
Comparison of frozen control and preserved bacterial DNA and RNA-TRFLP peak areas from laboratory samples preserved with BioDry.
Results shown for the comparison of pooled DNA samples (A, n = 3 per treatment) and average ribosomes of all replicates (B, n = 8 ± SD) from BioDried samples that were stored for T15 and T30 d (black and blue profiles respectively) with frozen samples (controls; n = 2). Values represent only those peaks present in more than one replicate for the treatments being compared; the number of OTUs used for each analysis is indicated at the bottom-right of each graph.
Fig 4.
Agarose gel of total nucleic acid extracts from seawater (A) and river water (B) field samples.
Samples were either flash frozen in liquid nitrogen and stored frozen (T0 Control) or preserved using the portable BioDry apparatus and stored for either T15 or T30 days at room temperature.
Table 3.
OTUs and Sorensen’s similarity index measure (CS) for the field verification tests.
The average number of bacterial, eukaryotic, and archaea OTUs (± SD) from the RNA-TRFLP profiles are indicated; (n = 3 for each treatment; na = not available).
Fig 5.
Comparison of RNA-TRFLP peak areas in frozen versus preserved seawater field samples.
Results are shown for seawater samples that were frozen, plotted against those preserved with BioDry and stored for either 10 or 30 d for the bacterial (A & B), eukaryotic (C & D), or archaeal (E & F) communities. Values represent averages (n = 3 ± SD) of only those peaks present in both controls and BioDried samples; the number of OTUs used for each analysis is indicated at the bottom-right of each graph.
Fig 6.
Comparison of RNA-TRFLP peak areas in frozen versus preserved river water field samples.
Results are shown for riverine samples that were frozen, plotted against those preserved with BioDry and stored for either 10 or 30 d for the bacterial (A & B) & eukaryotic (C & D) communities. Values represent averages (n = 3 ± SD) of only those peaks present in both controls and BioDried samples; the number of OTUs used for each analysis is indicated at the bottom-right of each graph.
Fig 7.
Assessment of protein and mRNA integrity for seawater field samples preserved with BioDry.
Gene expression for rbcL was assayed using clade-specific primers for diatoms (A) and haptophytes (B). Protein was assayed by western blot using an RbcL antibody (C). Red arrows represent assay positives for Emiliania huxleyi CCMP & Glycine max (soy) leaf extract respectively. Frozen controls (T0), preserved samples (T10 & T30 days), as well as the representative DNA/contamination controls are indicated.