Open AccessDeoxyribonuclease Treatment Improves the Homogeneity of Single-Stranded DNA Preparations
Author(s) -
Т. К. Алиев,
А. А. Панина,
В Г Коробко,
S.D. Varfolomeyev
Publication year - 1997
Publication title -
biotechniques/biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/97221st02
Subject(s) - deoxyribonuclease , dna , phagemid , capsid , microbiology and biotechnology , deoxyribonucleases , biology , bacteriophage , double stranded , transformation (genetics) , chemistry , biochemistry , virus , virology , gene , escherichia coli
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-tocircular 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.