conl12017-sup-0001-FigureS1.pdf638KFigure S1. General schematic of the eDNA – LTS species detection process. (1) Water samples are taken from aquatic environments (photo credit: Frank Oliver, Indiana DNR), (2) water sample is run through a filter, which captures eDNA fragments (photo credit: Univ. of Notre Dame), (3) eDNA is extracted directly from the filter paper, (4) PCR amplification of mtDNA fragment containing species diagnostic differences using universal primers, (5) hybridization of denatured, single-stranded PCR product with nanobeads covered with a short ∼30 bp oligonucleotide that will bind only to the target species, and (6) the diameter of the nanobeads in solution are measured by LTS platform, where a single peak around the original diameter of the beads will appear when no target species DNA is present and two peaks, one of the original size and one ∼150 bp larger, will appear when the target species is present. See Methods section for detailed descriptions of each step. Depicted in step 6 is the original (bench-top) implementation of LTS (Li et al. 2010, 2011), which has been modified in part to create a portable instrument, on which the data for these experiments were taken.
conl12017-sup-0002-TableS1.docx92KTable S1. Mean peak shift detected by LTS following hybridization for individual specimens and for the pooled total across all four specimens (all) for each of the five tested target species. Significance determined by one-sample t-tests to null hypothesis of no peak shift (= 0 nm). All means presented are least-square means of the four replicates, which remove the significant, but small effect of concentration (see Results).

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