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. 2016 Nov 28;45(5):e26. doi: 10.1093/nar/gkw1034

Figure 1.

Figure 1.

Concentrated Nicked DNA Circularization Assay (CNDCA) allows high yield formation of minicircle species. (A) Image of stained native polyacrylamide gel analyzing the differential formation of DNA minicircles and DNA multimers in a ligase-assisted circularization reaction with non-nicked (lanes 2–4) versus nicked (lanes 5–7) linear DNA substrates of 95 bp in size with an identical sequence. The circularization reactions were carried out at elevated DNA concentration (2 μM) in the presence and absence of the bending protein Abf2p as indicated. M denotes molecular weight DNA ladder and the size in bp is indicated on the left. Arrow and P indicate strand nick and phosphorylation at the 5΄ terminus, respectively. (B) Quantification of overall DNA multimers and minicircle species formed in the cyclization reaction in the presence of Abf2p with input nicked versus non-nicked DNA. (C) Electrophoretic mobility shift assay showing comparative binding activity of increasing concentration of the bending protein Abf2p with 95 bp linear nicked DNA used in CNDCA and nicked 95 bp minicircle of identical sequence.