Reduced viscosity polymer matrices for microchip electrophoresis of double-stranded DNA.
On a polymethylmethacrylate (PMMA) microchip, double-stranded DNA fragments with a wide size range from 50 bp to 20 kbp were separated by two polymer solutions. One was a hydroxypropylmethylcellulose-4000 (HPMC-4000) solution of 1.3% (w/v) to separate fragments below 590 bp, and another was a mixed four molecular weight poly(ethylene oxide) solution at a total concentration of 0.1% to separate fragments above 520 bp. The widths at half height (wh) of the fragments had a good relationship with their migration times (tR) in both polymer solutions. Such a relationship was suitable for obtaining the wh values of unresolved peaks, calculating the resolution of two adjacent fragments, and optimizing microchip separation matrices. Based on the relativity, a low viscosity medium containing 2% HPMC-50 and 8% glucose was optimized for high-performance separation of a phiX174 HaeIII restriction fragment digest.
Xu
Feng
F
Department of Medicinal Chemistry, Faculty of Pharmaceutical Sciences, The University of Tokushima, CREST, Japan Science and Technology, Shomachi, Tokushima 770-8505, Japan.
Jabasini
Mohammad
M
Liu
Shaoqian
S
Baba
Yoshinobu
Y
D004247:DNA / Q000032:analysis*; D004586:Electrophoresis / Q000379:methods; D006801:Humans; D008832:Microchemistry / Q000295:instrumentation; D011108:Polymers
D011108:Polymers; D004247:DNA