Figure-1d

Higher electrophoretic mobility DNA (HmDNA) in NaOH-denatured pUC19 is single-stranded in nature. pUC19 plasmid was denatured using 0.5 N NaOH to produce HmDNA. After ~5 min, NaOH was neutralized using 0.5 M HCl, and DNA was incubated with S1 nuclease, which would digest single-stranded DNA. HmDNA, but not native plasmid was degraded by S1 nuclease (lane 7). Experiment was repeated two times (i, ii).

Lane 1, 1 kb ladder; Lane 2, pUC19; Lane 3, pUC19 + 0.5 N NaOH; Lane 4, pUC19 + 0.5 N NaOH + 0.5 M HCl; Lane 5, pUC19 + S1 nuclease; Lane 6, pUC19 + 0.5 N NaOH + S1 nuclease; Lane 7, pUC19+ 0.5 N NaOH + 0.5 M HCl + S1 nuclease.