The current study analyzed the herpes simplex virus type 1 (HSV1) genome during suppressed and reactivated infection in vitro. Utilization of 3H-labeled HSV1 provided a highly specific probe for intracellular localization, isolation, and characterization of the HSV genome after infection of rabbit trigeminal ganglion neurons. Restriction enzyme analysis of viral DNA extracted during both suppressed and reactivated infection matched that of the HSV1 control DNA. Viral DNAs extracted from both cellular and nuclear fractions of host cells exhibited identical patterns. No detectable alterations in terminal fragments were observed, which suggests that the HSV1 genome is both linear and nonintegrated during suppressed infection in this system.