Regionally Based DNA Archives for Use As in Situ Validation for Edna Assays

EDNA is emerging as an effective tool for detecting the presence of DNA from cryptic or rare species without direct observation. A variety of methods for analyzing eDNA have also emerged. Foremost among these methods are conventional and quantitative PCR (qPCR) assays. Both methods require the amplification of short strands of DNA specific to target species. Validation of PCR assay specificity must be performed to ensure that cross reactivity with closely related and co-occurring species do not result in false positives. Typical validation of assay specificity includes in silico validation (BLAST) and when possible a cross reactivity panel using DNA from closely related and/or co-occurring species. While in-silico validation may take hours just assembling a clean cross reactivity DNA panel could take months. Locally and regionally sourced, vouchered, and sequenced DNA archives that are publically accessible would reduce duplicate research, decrease the use of assays validated in silico only, and aid in the expanded use of eDNA methods across regions. This presentation is intended to explore the idea of regional based DNA archives as a means to greatly reduce the time and effort required for PCR assay design and validation.