Deoxyribonuclease treatment improves the homogeneity of single-stranded DNA preparations.
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.
Aliev
T K
TK
Department of Chemical Enzymology, Chemical Faculty, Moscow State University, Russia. aliev@ibch.siobc.ras.ru
Panina
A A
AA
Korobko
V G
VG
Varfolomeyev
S D
SD
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
D004277:DNA, Single-Stranded; D004279:DNA, Viral; D003851:Deoxyribonucleases