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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...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Public Library of Science
2016
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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. |
format | Online Article Text |
id | pubmed-4972323 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
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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