Abstract
DNA replication in human melanoma cells is investigated by lysing the cells in dilute alkali. This lysis condition results in the release from parental DNA of the single-stranded DNA fragments located in active replicating units. The size of the released DNA should theoretically range from that of Okazaki-fragments up to that of the entire replication unit. However, the results showed that the released DNA replication intermediates which are detected range in the size between Okazaki-fragments up to 10 kb DNA fragments. The 10 kb DNA fragments show a discrete appearance in agarose gel electrophoresis. Moreover the kinetic results indicate that the ligation of the 10 kb DNA fragments to form high molecular weight DNA is a late step. A prerequisite for the release of this DNA fragment as a discrete population is that there are gaps in the continuity of the newly synthesized DNA spaced roughly 10 kb away from each other.
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