Fig. 6.

Double-stranded DNA synthesis in vitro. (A) Production of dsDNA depends on NTPs. Rolling-circle DNA replication was performed as described in figure legend 5, but in the presence of varying amounts of NTPs (0 μM, 37.5 μM, 75 μM, 150 μM, and 300 μM). Products were labeled with [α-³²P]dCTP, cleaved with MboI, separated by polyacrylamide gel electrophoresis, and detected by autoradiography. (B) Incomplete cleavage by MboI detects long double-stranded DNA products, indicative of rolling-circle DNA replication. The analysis was as in (A), but products labeled with either [α-³²P]dGTP (lagging strand) or [α-³²P]dCTP (leading strand) were cleaved with subsaturating amounts of MboI.