Ultra-fast high-resolution agarose electrophoresis of DNA and RNA using low-molarity conductive media

Jonathan R. Brody; Eric S. Calhoun; Eike Gallmeier; Talisa D. Creavalle; Scott E. Kern

doi:10.2144/04374st04

Ultra-fast high-resolution agarose electrophoresis of DNA and RNA using low-molarity conductive media

Jonathan R. Brody, Eric S. Calhoun, Eike Gallmeier, Talisa D. Creavalle, Scott E. Kern

Research output: Contribution to journal › Article › peer-review

17 Scopus citations

Abstract

Current DNA electrophoretic solutions employ high ionic concentrations and require long electrophoretic run times. Here we demonstrate that high and low molecular weight double-stranded DNA, single-stranded DNA (ssDNA), and RNA can be separated rapidly in agarose-based low-molarity conductive media. Separation of small DNA fragments was optimized by substituting 1-mM solutions of alkali metals or a nonbiological amine that distributed voltage with a minute current. These ultra-dilute solutions can separate DNA at least 15-fold faster. Low-molarity media at 5-10 mM adequately separated RNA and larger DNA fragments as well. These novel media reduce the Joule heating of the electrophoretic system and allow for easy-to-use, ultra-fast separation of DNA fragments.

Original language	English (US)
Pages (from-to)	598-602
Number of pages	5
Journal	BioTechniques
Volume	37
Issue number	4
DOIs	https://doi.org/10.2144/04374st04
State	Published - Oct 2004
Externally published	Yes

ASJC Scopus subject areas

Biotechnology
Biochemistry, Genetics and Molecular Biology(all)

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abstract = "Current DNA electrophoretic solutions employ high ionic concentrations and require long electrophoretic run times. Here we demonstrate that high and low molecular weight double-stranded DNA, single-stranded DNA (ssDNA), and RNA can be separated rapidly in agarose-based low-molarity conductive media. Separation of small DNA fragments was optimized by substituting 1-mM solutions of alkali metals or a nonbiological amine that distributed voltage with a minute current. These ultra-dilute solutions can separate DNA at least 15-fold faster. Low-molarity media at 5-10 mM adequately separated RNA and larger DNA fragments as well. These novel media reduce the Joule heating of the electrophoretic system and allow for easy-to-use, ultra-fast separation of DNA fragments.",

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AU - Gallmeier, Eike

AU - Creavalle, Talisa D.

AU - Kern, Scott E.

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AB - Current DNA electrophoretic solutions employ high ionic concentrations and require long electrophoretic run times. Here we demonstrate that high and low molecular weight double-stranded DNA, single-stranded DNA (ssDNA), and RNA can be separated rapidly in agarose-based low-molarity conductive media. Separation of small DNA fragments was optimized by substituting 1-mM solutions of alkali metals or a nonbiological amine that distributed voltage with a minute current. These ultra-dilute solutions can separate DNA at least 15-fold faster. Low-molarity media at 5-10 mM adequately separated RNA and larger DNA fragments as well. These novel media reduce the Joule heating of the electrophoretic system and allow for easy-to-use, ultra-fast separation of DNA fragments.

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Ultra-fast high-resolution agarose electrophoresis of DNA and RNA using low-molarity conductive media

Abstract

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