Design parameters of the quantitative PCR (qPCR) assay used to test nasal lavage samples obtained from three-toed box turtles (Terrapene carolina triunguis) for emydid mycoplasma, based on published 16s ribosomal RNA gene sequences of this microbe (Ossiboff et al. 2015). The calibration curve was based on three dilution curves, run in triplicate on three different plates. Although the assay had a detection limit of 50 copies, it was optimized for 500–50,000,000 copies of DNA.