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Evaluating non-detection risk associated with metabarcoding approaches for rare metabarcoding approaches
Citation:
Hatzenbuhler, C., J. Kelly, E. Pilgrim, AND J. Martinson. Evaluating non-detection risk associated with metabarcoding approaches for rare metabarcoding approaches. Presented at Upper Midwest Invasive Species Conference, Duluth, MN, October 20 - 23, 2014.
Impact/Purpose:
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Description:
Early detection of non-indigenous species (NIS), newly introduced species at low abundance in the monitoring area, can strengthen current management strategies including NIS control and eradication. A practical early detection strategy requires achieving balance between efficient sampling and taxonomy methods that minimize early detection costs such as time and effort with the risk associated with non-detection. For that reason, current technological advances in molecular taxonomy may prove useful for species identification regarding NIS early detection. The goal of our research is to evaluate the risk of non-detection associated with DNA based methods for species identification in complex biotic community samples (i.e., metabarcoding). Our study used larval fish and benthic invertebrates collected from the St. Louis River Estuary to construct independent and progressively complex experimental samples. By controlling sample composition and complexity we constructed samples to determine the lowest limit of detection (LLD) for the target (rare) species in each sample and investigate some factors that may affect detection limits. Next generation sequencing (NGS) was used to decode the mtDNA CO1 gene of each species present in a sample. Thus far our results have provided insight into NGS detection capabilities and logistical constraints. With this gained knowledge sample construction and experimental design can be modified to improve the probability of detection success. Recognizing the confines of DNA-based methods for species identification in this context allows for an accurate assessment of its potential use pertaining to the early detection of aquatic invasive species and implementation into a NIS monitoring program.