Detecting Jefferson salamander on-site using our point-of-need environmental DNA (eDNA) detection platform Background The Jefferson salamander (Ambystoma jeffersonianum) is characterized by a dark grey or brown back with lighter under-parts, with light bluish-grey flecks along the flanks and tail in some individuals (Fig 1). The species is distributed across northeastern U.S. and southern Ontario (Fig 2; MNRF), inhabiting deciduous or mixed upland forest with access to suitable breeding ponds. Breeding ponds are ephemeral or vernal, woodland pools that dry out in late summer/early fall. The species is in serious decline due to habitat loss, pollution of breeding ponds and disturbance to migratory routes. As such, the species is listed as Endangered (Species at Risk Ac

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