Ultra-fast high-resolution agarose electrophoresis of DNA and RNA using low-molarity conductive media.
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.
Brody
Jonathan R
JR
The Johns Hopkins University School of Medicine, Baltimore, MD 21231, USA. sk@jhmi.edu
Calhoun
Eric S
ES
Gallmeier
Eike
E
Creavalle
Talisa D
TD
Kern
Scott E
SE
D003470:Culture Media / Q000737:chemistry; D004247:DNA / Q000737:chemistry*; D004553:Electric Conductivity; D004587:Electrophoresis, Agar Gel / Q000295:instrumentation / Q000379:methods*; D005782:Gels; D008970:Molecular Weight; D012313:RNA / Q000737:chemistry*; D012996:Solutions; D013997:Time Factors
D003470:Culture Media; D005782:Gels; D012996:Solutions; D012313:RNA; D004247:DNA