Permeable cell systems have been developed by treatment with digitonin or saponin for studying in vitro DNA replication of chromatin. DNA replication of SV40 nucleoprotein complexes (SV40 chromatin) in these systems requires the addition of ATP, dCTP, dGTP, dTTP, and Mg2+. Further addition of three other ribonucleoside triphosphates slightly increased DNA synthesis. Omission of ATP greatly reduced DNA synthesis. In the presence of ATP, however, omission of dATP did not significantly alter or rather slightly enhanced DNA synthesis. Analysis of replicated SV40 DNA by gel electrophoresis and autoradiography indicated that complete replication of SV40 DNA and chromatin occurred in these systems