Modification of the electrokinetic properties of reversible electrophoresis gels for the separation and preparation of DNA - Addition of linear polymers
Kd. Cole et al., Modification of the electrokinetic properties of reversible electrophoresis gels for the separation and preparation of DNA - Addition of linear polymers, APPL BIOC B, 82(1), 1999, pp. 57-76
The effect of adding linear polymers to a novel reversible electrophoretic
media was measured. Reversible gels are formed using the polyanionic carboh
ydrate polymer, gellan gum. Gellan gum forms strong stable gels in the pres
ence of divalent cations or diamines. The gels are reversible (return to so
lution) by changing the ionic environment or pH. Gellan gum is an anionic p
olymer, and the electrophoresis gels have considerable electroosmotic flow
(EOF) toward the negative electrode. We measured the EOF in gellan gum elec
trophoresis gels as a function of gel concentration, buffer composition, an
d linear polymer additive. The linear polymers used in this study were poly
ethylene oxide and hydroxyethyl cellulose. Both polymers reduced EOF in the
gels, in a manner dependent on molecular weight. Polymers with a high mole
cular weight were more effective at reducing EOF. The addition of polymers
increased the resolution of low molecular weight DNA. Native gellan gum res
olved DNA from approx 50,000 to 1000 bp. Addition of the polymers resolved
DNA down to approx 50 bp, in some instances. The influence of the polymers
on circular plasmid DNA was also investigated. Addition of high molecular w
eight polyethylene oxide reduced the electrophoretic mobility of the nicked
circular form compared to the supercoiled form.