DNA capillary electrophoresis using poly(vinyl alcohol). II. Separation mediaWe represent the first extensive study on DNA capillary electrophoresis using various poly(vinyl alcohol) (PVAL)-related polymers as separation media. As a separation medium, PVAL homopolymer has shown poor resolutions probably due to its very strong hydrogen-bonding characteristics, resulting in extensive self-aggregation. On the other hand, poly(vinyl alcohol-co-vinyl acetate) and poly(vinyl alcohol-co-1-vinyl-2-pyrrolidone) (PVAL-VP), both with a degree of polymerization of ,3.0610 3 , have been found to give excellent electropherograms with good resolutions for the analysis of double-stranded (ds)DNA. PVAL-VP, with hydrolytic stability in high and low pH regions, has also yielded fair electropherograms for single-stranded (ss)DNA under neutral and alkaline conditions, although further investigation is essential in order to increase the resolutions necessary for DNA sequencing analysis. The separations obtained under alkaline conditions have shown significantly shorter retention times, one-third of that for the current commercial separation media, due to the higher ionization of phosphate groups in the DNA chains.
IntroductionDuring the recent marked advances in capillary electrophoresis (CE), a number of water-soluble polymers have been investigated as separation media: poly(N,N'-dimethylacrylamide) (PDMA), poly(acrylamide) (PAM), agarose, hydroxyethylcellulose (HEC), polyethylene glycol (PEG), poly(vinylpyrrolidone) (PVP), pluronic polyols, etc. [1][2][3][4]. However, only two papers have been published on the application of poly(vinyl alcohol) (PVAL) in CE, in spite of its apparent suitability as a typical nonionic water-soluble synthetic polymer. Simò-Alfonso et al. [5] have reported successful results for proteins using PVAL (M r 133 000), which offers excellent resolutions in 4-6 wt% solutions and shows sieving effects even in extremely dilute solutions, with concentrations of ,1 wt%, that is much lower than the observed threshold of 3 wt% from the viscosity data. Kleemiss et al. [6] have shown deterioration of resolution about the CE of double-stranded (ds)DNA using the PVAL of M r 50 000 and hydrolysis grade 99%, with bare fused-silica capillaries. They ascribed the negative results to strong self-aggregation of PVAL by hydrogen bonding. Both previous works have attributed the specific characteristics of PVAL to hydrogen bonding, which should be controlled in the application to CE.PVAL is believed to have a significant potential in CE because it has an advantage of the hydrolytic stability all over pH ranges, together with additional advantages, such as moderate viscosities, environmental friendliness of nontoxic and biodegradable properties, and full transparency in the UV region down to 200 nm [5]. The intrinsic chemical stability of PVAL is useful for expanding applications of CE to higher and lower pH regions for the separations and analyses of DNA fragments, proteins, and other charged molecules. In particular, alkaline...