title: Deoxyribonuclease treatment improves the homogeneity of single-stranded DNA preparations.
abstract: The isolation of single-stranded (ss) phagemid DNA using standard protocols often results in impure preparations, which contain undesirable quantities of chromosomal and/or double-stranded (ds) phagemid DNA. Here we report a simple and efficient method for elimination of virtually all dsDNA by incubation of phagemid viral particles with deoxyribonuclease I. In addition to analyzing the ratio of linear-to-circular topological forms of ssDNA after deoxyribonuclease I treatment, we verified that no decrease in transformation efficiency occurred and demonstrated that ssDNA molecules covered by capsid proteins remained intact following such treatment.
authors.affiliation: Department of Chemical Enzymology, Chemical Faculty, Moscow State University, Russia. aliev@ibch.siobc.ras.ru
authors.forename: T K
authors.initials: TK
authors.lastname: Aliev
authors.forename: A A
authors.initials: AA
authors.lastname: Panina
authors.forename: V G
authors.initials: VG
authors.lastname: Korobko
authors.forename: S D
authors.initials: SD
authors.lastname: Varfolomeyev
mesh_terms: D001435:Bacteriophages / Q000235:genetics; D004277:DNA, Single-Stranded / Q000302:isolation & purification*; D004279:DNA, Viral / Q000302:isolation & purification*; D003851:Deoxyribonucleases / Q000737:chemistry*; D004587:Electrophoresis, Agar Gel; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D009691:Nucleic Acid Denaturation; D010957:Plasmids; D014170:Transformation, Genetic
chemical_list: D004277:DNA, Single-Stranded; D004279:DNA, Viral; D003851:Deoxyribonucleases