Isolation of in vitro synthesized covalently closed circular double-stranded DNA by selective denaturation and filtration.
Based upon their resistance to irreversible denaturation, covalently closed circular (CCC) DNA (non-nicked, double-stranded circular molecules) can be purified by alkaline denaturation, neutralization, and filtration through a nitrocellulose membrane. This procedure offers a simple means of isolating in vitro synthesized CCC DNA molecules. The preparations of molecules obtained by this method consisted of 91-97% CCC DNA and contained no detectable inhibitors of biological activity or enzymatic digestion.
D010584:Bacteriophage phi X 174 / Q000235:genetics; D001435:Bacteriophages / Q000235:genetics*; D003101:Collodion; D004270:DNA, Circular / Q000096:biosynthesis / Q000302:isolation & purification*; D004279:DNA, Viral / Q000096:biosynthesis / Q000302:isolation & purification*; D004587:Electrophoresis, Agar Gel; D004926:Escherichia coli; D005374:Filtration; D009691:Nucleic Acid Denaturation; D009695:Nucleic Acid Renaturation; D009838:Oligodeoxyribonucleotides / Q000235:genetics
D004270:DNA, Circular; D004279:DNA, Viral; D009838:Oligodeoxyribonucleotides; D003101:Collodion