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The Miraprep: A Protocol that Uses a Miniprep Kit and Provides Maxiprep Yields

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 th...

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Detalles Bibliográficos
Autores principales: Pronobis, Mira I., Deuitch, Natalie, Peifer, Mark
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4972323/
https://www.ncbi.nlm.nih.gov/pubmed/27486871
http://dx.doi.org/10.1371/journal.pone.0160509
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author Pronobis, Mira I.
Deuitch, Natalie
Peifer, Mark
author_facet Pronobis, Mira I.
Deuitch, Natalie
Peifer, Mark
author_sort Pronobis, Mira I.
collection PubMed
description 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.
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spelling pubmed-49723232016-08-18 The Miraprep: A Protocol that Uses a Miniprep Kit and Provides Maxiprep Yields Pronobis, Mira I. Deuitch, Natalie Peifer, Mark PLoS One Research Article 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. Public Library of Science 2016-08-03 /pmc/articles/PMC4972323/ /pubmed/27486871 http://dx.doi.org/10.1371/journal.pone.0160509 Text en © 2016 Pronobis et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Pronobis, Mira I.
Deuitch, Natalie
Peifer, Mark
The Miraprep: A Protocol that Uses a Miniprep Kit and Provides Maxiprep Yields
title The Miraprep: A Protocol that Uses a Miniprep Kit and Provides Maxiprep Yields
title_full The Miraprep: A Protocol that Uses a Miniprep Kit and Provides Maxiprep Yields
title_fullStr The Miraprep: A Protocol that Uses a Miniprep Kit and Provides Maxiprep Yields
title_full_unstemmed The Miraprep: A Protocol that Uses a Miniprep Kit and Provides Maxiprep Yields
title_short The Miraprep: A Protocol that Uses a Miniprep Kit and Provides Maxiprep Yields
title_sort miraprep: a protocol that uses a miniprep kit and provides maxiprep yields
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4972323/
https://www.ncbi.nlm.nih.gov/pubmed/27486871
http://dx.doi.org/10.1371/journal.pone.0160509
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