The temperature of the isolation medium exerted no significant effect on the rate of positive cultures for Neisseria gonorrhoeae. However, significantly more colonies were obtained on ambient-temperature medium than on cold (4C) medium. A small number of specimens (six) failed to grow on cold medium but grew on ambient-temperature medium. Viable counts showed essentially no effect of medium temperature on the nondiscriminating strain, a 90% reduction on the cold medium for the cold-intolerant strain, and for the strains with warm medium preference, about twice as many colonies on warm as on cold or ambient-temperature medium.