The Black-spotted Newt (Notophthalmus meridionalis) is a chronically understudied species, with much information on the natural history, ecology, and distribution of this species poorly known. Previous studies using traditional methodologies have had limited success detecting N. meridionalis. A novel eDNA assay was designed and implemented with the goal of better understanding N. meridionalis distribution in south Texas. Primers were designed to amplify N. meridionalis DNA and tested against DNA from sympatric amphibians. This assay combined with traditional methods was implemented at 62 sites throughout south Texas, and N. meridionalis was detected at eight localities. At each site surveyed, soil, water, and habitat data were recorded, and these were tested with a logistic regression against N. meridionalis presence/absence. The most parsimonious models showed that paved road absence, ephemerality, higher soil copper levels, lower soil sand percentage, and lower water conductivity were associated with N. meridionalis presence
