DNA replication in human fibroblasts with normal excision repair was investigated after ultraviolet irradiation and incubation with caffeine or hydroxyurea. The DNA synthesized soon after irradiation had a reduced size, but that synthesized later was near normal size. When caffeine was present before labeling, it reduced the size of DNA synthesized but when added after labeling it was without effect. When irradiated cells were allowed to grow, labeled DNA increased in size steadily for 60 min to a maximum that was below control and dose-dependent. Further growth resulted in a transition of some label to parental DNA sized, but a large fraction remained permanently blocked at smaller sizes, producing bimodal distributions of DNA. The steady increase in size was inhibited by hydroxyurea. Removing cells from hydroxyurea resulted in increases similar to or slightly slower than those observed immediately after labeling, and this protocol did not permit cells to acquire any induced or enhanced capacity to replicate damaged DNA.