Separation of double‐stranded DNA fragments by capillary electrophoresis: Impacts of poly(ethylene oxide), gold nanoparticles, ethidium bromide, and pH

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.

Electrophoretic Separation of Proteins in Capillaries Filled with Poly(ethylene oxide)-stabilized Silver Nanoparticles

“…14 Chang's group employed gold nanoparticles (GNPs) as additives in CE separation of DNA fragments. [23][24][25][26] Due to the interaction of DNA fragments with the GNPs, the apparent mobilities of DNA fragments changed, and the plate numbers doubled.…”

Capillary electrophoretic separation of biologically active amines and acids using nanoparticle‐coated capillaries

“…Different NPs have also been used in CE for improved separation. Gold NPs which are easily prepared were separately added to aqueous electrolytes and poly (ethylene oxide) solutions to optimize resolution for small solutes, as well as DNA and proteins [24][25][26][27][28][29][30][31]. Continuous filling and partial filling of polymeric NPs that serve as pseudostationary phases are both useful for the separations of three amines by CEC, without conducting time-consuming particle packing and using retaining frits [32].…”

Effect of Nanoparticles in Protein Separation by Capillary Electrophoresis

“…To overcome these difficulties, the developments on the separation matrix have focused on the combination of good separation efficiency with dynamic coating and low viscosity for high throughput analysis. For example, mannitol added poly-N-isopropylacrylamide (PNIPAM), 23 and gold nanoparticle (GNP) 24,25 showed the potential for this purpose. However, it turned out that mannitoladded PNIPAM had a little dynamic coating ability for the reproducible DNA separation.…”