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

被引:44
作者
Brody, JR [1 ]
Calhoun, ES [1 ]
Gallmeier, E [1 ]
Creavalle, TD [1 ]
Kern, SE [1 ]
机构
[1] Johns Hopkins Univ, Sidney Comprehens Canc Ctr, Dept Oncol, Sch Med, Baltimore, MD 21231 USA
关键词
D O I
10.2144/04374ST04
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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 arnine 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.
引用
收藏
页码:598 / +
页数:3
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