Analysis of double-stranded DNA by microchip capillary electrophoresis using polymer solutions containing gold nanoparticles

Yang-Wei Lin, May Jen Huang, Huan Tsung Chang

Research output: Contribution to journalArticle

55 Citations (Scopus)

Abstract

The impact of gold nanoparticles (GNPs) on the microchip electrophoretic separation of double-stranded (ds) DNA using poly(ethylene oxide) (PEO) is described. Coating of the 75-μm separation channel on a poly(methyl methacrylate) (PMMA) plate in sequence with poly(vinyl pyrrolidone), PEO, and 13-nm GNPs is effective to improve reproducibility and resolution. In this study, we have also found that adding 13-nm GNPs to 1.5% PEO is extremely important to achieve high resolution and reproducibility for DNA separation. In terms of the stability of the GNPs, 100 mM glycine-citrate buffer at pH 9.2 is a good buffer system for preparing 1.5% PEO. The separation of DNA markers V and VI ranging in size from 8 to 2176 base pairs has been demonstrated using the three-layer-coated PMMA microdevice filled with 1.5% PEO containing the GNPs. Using these conditions, the analysis of the polymerase chain reaction products of UGT1A7 was complete in 7 min, with the relative standard deviation values of the peak heights and migration times less than 2.3% and 2.0%, respectively. In conjunction with stepwise changes of the concentrations of ethidium bromide (0.5 and 5 μg/ml), this method allows improved resolution and sensitivity for DNA markers V and VI.

Original languageEnglish
Pages (from-to)47-55
Number of pages9
JournalJournal of Chromatography A
Volume1014
Issue number1-2
DOIs
Publication statusPublished - 2003 Oct 3

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Microchip Electrophoresis
Capillary electrophoresis
Capillary Electrophoresis
Polymer solutions
Polyethylene oxides
Oligonucleotide Array Sequence Analysis
Gold
Nanoparticles
Polymers
DNA
Polymethyl Methacrylate
Genetic Markers
Buffers
Pyrrolidinones
Ethylene Oxide
Ethidium
Citric Acid
Base Pairing
Polymerase chain reaction
Glycine

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Biochemistry
  • Organic Chemistry

Cite this

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abstract = "The impact of gold nanoparticles (GNPs) on the microchip electrophoretic separation of double-stranded (ds) DNA using poly(ethylene oxide) (PEO) is described. Coating of the 75-μm separation channel on a poly(methyl methacrylate) (PMMA) plate in sequence with poly(vinyl pyrrolidone), PEO, and 13-nm GNPs is effective to improve reproducibility and resolution. In this study, we have also found that adding 13-nm GNPs to 1.5{\%} PEO is extremely important to achieve high resolution and reproducibility for DNA separation. In terms of the stability of the GNPs, 100 mM glycine-citrate buffer at pH 9.2 is a good buffer system for preparing 1.5{\%} PEO. The separation of DNA markers V and VI ranging in size from 8 to 2176 base pairs has been demonstrated using the three-layer-coated PMMA microdevice filled with 1.5{\%} PEO containing the GNPs. Using these conditions, the analysis of the polymerase chain reaction products of UGT1A7 was complete in 7 min, with the relative standard deviation values of the peak heights and migration times less than 2.3{\%} and 2.0{\%}, respectively. In conjunction with stepwise changes of the concentrations of ethidium bromide (0.5 and 5 μg/ml), this method allows improved resolution and sensitivity for DNA markers V and VI.",
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Analysis of double-stranded DNA by microchip capillary electrophoresis using polymer solutions containing gold nanoparticles. / Lin, Yang-Wei; Huang, May Jen; Chang, Huan Tsung.

In: Journal of Chromatography A, Vol. 1014, No. 1-2, 03.10.2003, p. 47-55.

Research output: Contribution to journalArticle

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