Biotechniques. 2004 Oct;37(4):598, 600, 602.

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

Brody JR, Calhoun ES, Gallmeier E, Creavalle TD, Kern SE.

Source

The Johns Hopkins University School of Medicine, Baltimore, MD 21231, USA. sk@jhmi.edu

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.

PMID:: 15517972; [PubMed - indexed for MEDLINE]

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Research Support, U.S. Gov't, P.H.S.

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CA62924/CA/NCI NIH HHS/United States

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