qPCR results from design and partial validation of three novel eDNA qPCR assays for several common North American tick (Arachnida: Ixodida) species

The range expansion of ticks to higher latitudes poses a severe threat to human health exposing human populations who had no prior contact with ticks to several harmful tick-borne diseases.  Early detection of ticks in new areas is critical to help inform the public and medical professionals of the dangers associated with tick encounters.  Environmental DNA represents a novel survey method that could provide reliable records of tick occurrences and timely warnings of their range expansions.  In this study, we designed three novel eDNA qPCR assays for three common North American tick species (Dermacentor variabilis, Amblyomma americanum, and Ixodes scapularis) and tested them on samples of grasses collected from grasslands and forests in Illinois.  We provide in silico and in vitro validation of all three assays, however we were unable to generate any positive detections from field samples.  Our lack of eDNA detections likely stems from low eDNA deposition rates coupled with rapid degrad..., Plant matter samples containing grasses and leaf litter were collected in the fall of 2020, and from May through October of 2021 from various sites throughout central and southern Illinois.  Three 250 meter linear transects were established at each site and sampled for ticks and leaf litter via random selection (without replacement) to reduce bias (i.e. one of the three transects was selected at random for each site per visit, for a total of three visits on average).  50mL Falcon tubes were filled halfway with unpacked plant matter collected at approximately 0m, 150m, and 250m along the transect to ensure adequate coverage. The leaf litter sample tube and a control (blank) 50 mL tube were filled with enough CTAB extraction buffer to cover all plant matter, and tightly capped. Both tubes were inverted five-ten times (as necessary) to ensure all litter was in contact with the buffer.
All field samples were extracted 30 days post-collection via a modified chloroform: isoamyl alcohol method..., , # Tick qPCR Results

This file contains 2 datasets. The first contains all of the Cq values from the qPCR plates used to develop limit of detection (LOD) and limit of quantitation (LOQ) values for 3 qPCR assays targeting 3 species of ticks in Illinois (*Dermacentor variabilis*, *Amblyomma americanum*, and *Ixodes scapularis*). The second dataset contains qPCR results from the same three qPCR assays on eDNA samples, collected from grasslands and forests in Illinois.

## Description of the data and file structure

The first dataset qPCR results from 3 plates that were used to develop LOD and LOQ values for each of the assays. To calculate these values for each assay, we ran qPCR plates containing 8 standards with 12 different replicates per standard. Each standard contained a known concentration of DNA from a gBlock ranging from 1 copy/reaction to 3*10^5 copies/reaction. The resulting Cq values were then used to calculate the LOD and LOQ according to the procedure outlined in Klymus...