title: The Miraprep: A Protocol that Uses a Miniprep Kit and Provides Maxiprep Yields.
abstract: Plasmid purification is a basic tool of molecular biologists. Although the development of plasmid isolation kits utilizing silica spin columns reduced the time and labor spent on plasmid purification, achieving large plasmid DNA yields still requires significant time and effort. Here we introduce the Miraprep, a rapid protocol that allows isolation of plasmid DNA using commercial Miniprep kits, but with DNA yields comparable to commercial Maxiprep plasmid purifications. Combining ethanol precipitation with spin column purification, we created a DNA isolation protocol that yields highly concentrated plasmid DNA samples in less than 30 minutes. We show that Miraprep isolated plasmids are as stable as plasmids isolated by standard procedures, can be used for standard molecular biology procedures including DNA sequencing, and can be efficiently transfected into mammalian cells. This new plasmid DNA isolation protocol will significantly reduce time and labor without increasing costs.
authors.affiliation: Curriculum in Genetics and Molecular Biology, University of North Carolina at Chapel Hill, Chapel Hill, NC, United States of America.
authors.forename: Mira I
authors.initials: MI
authors.lastname: Pronobis
authors.affiliation: Department of Biology, University of North Carolina at Chapel Hill, CB#3280, Chapel Hill, NC, United States of America.
authors.forename: Natalie
authors.initials: N
authors.lastname: Deuitch
authors.affiliation: Curriculum in Genetics and Molecular Biology, University of North Carolina at Chapel Hill, Chapel Hill, NC, United States of America.|Department of Biology, University of North Carolina at Chapel Hill, CB#3280, Chapel Hill, NC, United States of America.
authors.forename: Mark
authors.identifier: http://orcid.org/0000-0003-1412-3987
authors.initials: M
authors.lastname: Peifer
mesh_terms: D001431:Bacteriological Techniques / Q000379:methods; D002460:Cell Line; D003001:Cloning, Molecular / Q000379:methods*; D003132:Commerce; D004247:DNA / Q000302:isolation & purification*; D004269:DNA, Bacterial / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D010957:Plasmids / Q000302:isolation & purification*; D011506:Proteins / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000302:isolation & purification*; D014162:Transfection / Q000379:methods
chemical_list: D004269:DNA, Bacterial; D011506:Proteins; D011994:Recombinant Proteins; D004247:DNA