Abstract.—The smallmouth salamander (Ambystoma texanum) is a widely distributed member of the family Ambystomatidae found throughout much of the central United States. We developed primers targeting a 147 base pair fragment of the mitochondrial cytochrome b region of the smallmouth salamander and tested these primers in silico, in vitro, and in vivo in laboratory-based studies. We believe these validated tools will be widely useful in the detection of A. texanum in breeding ponds, particularly in habitats where multiple species of Ambystoma occur.

Key Words.—conservation, eDNA, salamander, Ambystomatidae, Ambystoma texanum