Deoxyribonuclease treatment improves the homogeneity of single-stranded DNA preparations |
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Authors: | TK Aliev AA Panina VG Korobko SD Varfolomeyev |
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Affiliation: | Department of Chemical Enzymology, Chemical Faculty, Moscow State University, Russia. aliev@ibch.siobc.ras.ru |
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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. |
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