CE of dsDNA in low-molecular-weight polyethylene oxide solutions.
To realise effective size separations of nucleic acid fragments using CE, gel-based matrices are commonly employed. The separation of label-free dsDNA ladders and plasmid fragments in an uncross-linked semi-dilute poly (ethylene) oxide solution using multi-pixel UV detection at 254 nm is reported. Improvements in the sensitivity of UV detection of dsDNA using signal averaging over multiple pixels is demonstrated. Separations performed using a diode array detector also allow the progress of the separation to be monitored as a function of time. Several polymers were examined including methyl cellulose, linear polyacrylamide, hydroxy (propyl) methylcellulose and polyethylene oxide. Operations parameters investigated included UV transparency, self-coating capacity and separation efficiency. The results show complete resolution of all fragments under a range of conditions, including short separation lengths.
Pereira
Fiona
F
Department of Chemistry, Imperial College London, Exhibition Road, South Kensington, London, UK.
Hassard
Stuart
S
Hassard
John
J
deMello
Andrew
A
D004247:DNA / Q000032:analysis*; D019075:Electrophoresis, Capillary / Q000379:methods*; D008970:Molecular Weight; D011092:Polyethylene Glycols / Q000737:chemistry*; D012680:Sensitivity and Specificity; D012996:Solutions; D013056:Spectrophotometry, Ultraviolet; D013696:Temperature; D014783:Viscosity
D012996:Solutions; D011092:Polyethylene Glycols; D004247:DNA