Fig. 6. Abundance of nascent DNA within the 0.5-1 kb and 1-2 kb DNA fractions at three origins in HCT116. Nascent DNA from an asynchronous culture of HCT116, prepared as described in Materials and Methods, was fractionated on a nondenaturing 1% agarose gel following heat denaturation. Regions containing DNA spanning 0.5-1 kb and 1-2 kb were cut out and purified. Abundance of nascent DNA within those fractions was determined using realtime PCR, at three origins: lamin B2 (A), ß-globin (B) and c-myc (C) origins. As described in Fig. 5, LB2, BG40.9 and Myc11 are the origin-containing regions for these origins, whereas LB2C1, BG72 and Myc1 are distal origin-lacking regions lacking origin activity. The average of three experiments and 1SD is indicated. An asterisk (^*) represents a statistically significant difference (P<0.05) between the 0.5-1 kb and 1-2 kb DNA fractions in the abundance of nascent DNA at the specified DNA regions.