Thermotaxis (TTX) is one of the sophisticated behaviors in the nematode Caenorhabditis elegans. Although the mechanisms of thermotaxis have been deduced from different studies, they are controversial. Previous studies proposed a behavioral model where thermotaxis is regulated by the counterbalance between two opposite driving forces, while recent studies proposed stochastic models. In this study, we analyzed thermotaxis by a novel quantitative population TTX assay using a gentle linear thermal gradient. Analysis of thermotaxis in wild type animals revealed a clear thermal preference to a cultivation temperature with regard to the distribution of animals and the TTX mean expressing temperature preference. A time course assay revealed that the behavioral response to a preferred temperature was initially suppressed for at least 15 min in the animals cultivated at 23 degrees C, but not in those cultivated at 17 degrees C. Our result provides a possible explanation for the inconsistency between the various studies on thermotaxis and is consistent with the early behavioral model, where thermotaxis is regulated by the counterbalance between two driving forces.