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Improved separation of double‐stranded DNA fragments by capillary electrophoresis using poly(ethylene oxide) solution containing colloids
Author(s) -
Huang MingFeng,
Huang ChihChing,
Chang HuanTsung
Publication year - 2003
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200305432
Subject(s) - capillary electrophoresis , ethylene oxide , polyvinylpyrrolidone , capillary action , electrophoresis , chemistry , resolution (logic) , chromatography , colloidal gold , dna , analytical chemistry (journal) , nanoparticle , materials science , polymer , nanotechnology , polymer chemistry , organic chemistry , copolymer , biochemistry , artificial intelligence , computer science , composite material
The analysis of double‐stranded (ds) DNA fragments by capillary electrophoresis (CE) using poly(ethylene oxide) (PEO) solution containing gold nanoparticles (GNPs) is presented, focusing on evaluating size dependence of the GNPs and PEO on resolution and speed. To prevent the interaction of the capillary wall with DNA, the capillary was dynamically coated with polyvinylpyrrolidone. Using different PEO solutions containing GNPs ranging in diameter from 3.5 to 56 nm, we have achieved reproducible, rapid, and high‐resolution DNA separations. The results indicate that the sizes of PEO and GNPs as well as the concentration of PEO affect resolution. The separation of DNA ranging in size from 8 to 2176 base pairs (bp) was accomplished in 5 min using 0.2% PEO (8 MDa) containing 56 nm GNPs. We have also demonstrated the separations of the DNA fragments ranging from 5 to 40 kbp using 0.05% PEO (2 MDa) containing 13 nm GNPs or 0.05% PEO (4 MDa) containing 32 nm GNPs. With very low viscosity (< 15 cP), automatic replacement of the sieving matrices is easy, indicating a great potential for high‐throughput DNA analysis using capillary array electrophoresis systems.