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Separation of double-stranded DNA fragments by capillary electrophoresis: Impacts of poly(ethylene oxide), gold nanoparticles, ethidium bromide, and pH
- Source :
- ELECTROPHORESIS. 25:2186-2192
- Publication Year :
- 2004
- Publisher :
- Wiley, 2004.
-
Abstract
- The separation of DNA by capillary electrophoresis using poly(ethylene oxide) (PEO) containing gold nanoparticles (GNPs) is presented. The impacts of PEO, GNPs, ethidium bromide (EtBr), and pH on the separation of double-stranded DNA have been carefully explored. Using a capillary dynamically coated with 5.0% poly(vinylpyrrolidone) and filled with 0.2% PEO containing 0.3 x GNPs (the viscosity less than 15 cP), we have demonstrated the separation of DNA markers V and VI within 5 min at pH 8.0 and 9.0. In terms of resolution and reproducibility, GNPs have a greater impact on the separation of DNA at pH 9.0. Resolution improvements for large DNA fragments (> 300 base pairs, bp) are greater than those for small ones in the presence of GNPs. It is important to point out that reproducibility is excellent (relative standard deviations for the migration times less than 0.5%) and thus no further dynamic coating is required in at least 20 consecutive runs in the presence of GNPs. Using 0.2% PEO (pH 9.0) containing 0.3 x GNPs, the separation of DNA fragments ranging in size from 21 to 23,130 bp was accomplished in 7 min. The results presented in this study show the advantage of PEO containing GNPs for DNA separation, including rapidity, high resolving power, excellent reproducibility, and ease of filling capillaries.
- Subjects :
- Chromatography
Ethylene oxide
Capillary action
Base pair
Clinical Biochemistry
Oxide
Electrophoresis, Capillary
DNA
Hydrogen-Ion Concentration
Biochemistry
Nanostructures
Polyethylene Glycols
Analytical Chemistry
chemistry.chemical_compound
Capillary electrophoresis
chemistry
Colloidal gold
Ethidium
Gold
Ethidium bromide
Subjects
Details
- ISSN :
- 15222683 and 01730835
- Volume :
- 25
- Database :
- OpenAIRE
- Journal :
- ELECTROPHORESIS
- Accession number :
- edsair.doi.dedup.....35572c58178d21905bc9fae2f584b510