CE of dsDNA in low-molecular-weight polyethylene oxide solutions

被引:5
|
作者
Pereira, Fiona [1 ]
Hassard, Stuart [2 ]
Hassard, John [2 ]
deMello, Andrew [1 ]
机构
[1] Univ London Imperial Coll Sci Technol & Med, Dept Chem, London SW7 2AZ, England
[2] DeltaDOT Ltd, London, England
关键词
CE; DNA; Poly (ethylene) oxide; CAPILLARY GEL-ELECTROPHORESIS; POLYMERASE-CHAIN-REACTION; SINGLE-STRANDED-DNA; REPLACEABLE LINEAR POLYACRYLAMIDE; POLY(ETHYLENE OXIDE); MIGRATION BEHAVIOR; HYDROXYPROPYLMETHYL CELLULOSE; FLUORESCENCE DETECTION; ELECTROOSMOTIC FLOW; SIEVING MATRIX;
D O I
10.1002/elps.200900144
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
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页码:2100 / 2109
页数:10
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