TABLE 2.

Approximate cost analysis for processing one sample, including filtering, eDNA extraction, library preparation, and MiSeq sequencing at the Genomics Facility of the Advanced Analysis Centre (AAC) at the University of Guelph

Laboratory steps for processing one sample	Cost per step CAD/USD b (this study)	Cost per step CAD/USD b (Illumina protocol)
Filters (pore size 1 μm, diameter 47 mm; an average of 2.5 per trap)	2.5	2.5
CTAB extraction method (4 per filter)	2.5	2.5
DNA quantification (Qubit) and normalization	2.5	2.5
First PCR (pooled extracts per filter)	5	5
Magnetic beads (at all stages)	0.7	7
Second PCR (Indexing reaction and primers a )	7	25
HTS Sequencing (Illumina MiSeq)	30	30
Overall	50.2 CAD/36.7 USD	74.5 CAD/54.5 USD

Processing steps are compared to library preparation costs of Illumina's 16S Metagenomic Sequencing Library Preparation (“16S Metagenomic Sequencing Library Preparation”, 2013). Shaded cells represent steps common to both protocols where modifications were completed to reduce the protocol's total cost.

Nonhighlighted fields represent steps common to both protocols where no modifications were needed. The costs apply to all three molecular markers: COI, 16S, and ITS.

^aIllumina protocol adapted by the AAC at the University of Guelph.

^bCurrency exchange rates (CAD/USD) as of 09 August 2020:1 CAD = 0.73105.