Capillary electrophoresis of DNA in entangled polymer solutions

Grossman, Paul D.; Soane, David S.

doi:10.1016/0021-9673(91)80076-s

Cited by 158 publications

(59 citation statements)

References 17 publications

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“…As a second method, the Electrophoresis 1999, 20, 1962±1977 Separation of double-stranded and single-stranded DNA 1965 Table 1. Free mobility of ssDNA and dsDNA measured in capillary electrophoresis with 0% pDMA at different temperature with and without urea threshold was also determined by measuring the viscosity of the polymer solution at different concentrations and finding the point of departure from linearity on the double logarithmic specific viscosity vs. concentration plot (not shown) [31]. Also, by using the independently measured intrinsic viscosity values, we have previously determined the corresponding c* values of the different pDMA preparations (see Table 1 in [8]).…”

Section: Characterization Of the Separation Matrixmentioning

confidence: 99%

Separation of double-stranded and single-stranded DNA in polymer solutions: I. Mobility and separation mechanism

Heller

1999

Electrophoresis

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We have studied the separation of single‐stranded and double‐stranded DNA in a matrix of entangled, linear poly‐N,N‐dimethylacrylamide. Our results give better insight into the mechanisms involved during separations in polymer solutions. The dependence of different parameters on DNA size, electric field, pore size and the polymer chain length are evaluated and compared to theoretical predictions. Striking differences between experimental data and predicted scaling laws are found. Our data should help to optimize DNA separation in capillary electrophoresis and to improve existing models for DNA separation in porous matrices.

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Section: Characterization Of the Separation Matrixmentioning

confidence: 99%

Separation of double-stranded and single-stranded DNA in polymer solutions: I. Mobility and separation mechanism

Heller

1999

Electrophoresis

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“…The preferred mechanism of separation for short fragments follows the Ogston model. 23 In this case, the corresponding point of 100 bp was eliminated for the calculation of correlation coefficient (r 2 ), obtaining a value of 0.9974 for the region ranging from 200 to 1000 bp. The data are in agreement with those reported previously by our group, which found r 2 ¼ 0.9948 for fragments between 201 and 2036 bp for separations carried out on silica capillaries and HEC 140-160 kDa as separation matrix.…”

Section: Linearity Of Dna Sizingmentioning

confidence: 99%

Disposable polyester–toner electrophoresis microchips for DNA analysis

Duarte

Coltro

Borba

et al. 2012

Analyst

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Microchip electrophoresis has become a powerful tool for DNA separation, offering all of the advantages typically associated with miniaturized techniques: high speed, high resolution, ease of automation, and great versatility for both routine and research applications. Various substrate materials have been used to produce microchips for DNA separations, including conventional (glass, silicon, and quartz) and alternative (polymers) platforms. In this study, we perform DNA separation in a simple and low-cost polyester-toner (PeT)-based electrophoresis microchip. PeT devices were fabricated by a directprinting process using a 600 dpi-resolution laser printer. DNA separations were performed on PeT chip with channels filled with polymer solutions (0.5% m/v hydroxyethylcellulose or hydroxypropylcellulose) at electric fields ranging from 100 to 300 V cm À1 . Separation of DNA fragments between 100 and 1000 bp, with good correlation of the size of DNA fragments and mobility, was achieved in this system. Although the mobility increased with increasing electric field, separations showed the same profile regardless of the electric field. The system provided good separation efficiency (215 000 plates per m for the 500 bp fragment) and the separation was completed in 4 min for 1000 bp fragment ladder. The cost of a given chip is approximately $0.15 and it takes less than 10 minutes to prepare a single device.

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“…7 According to the Ogston theory, the DNA fragment keeps a spherical and rigid conformation of radius Rg. This particle moves across a random net formed by the gel fibers.…”

Section: Dna Migration Mechanisms In Polymeric Matricesmentioning

confidence: 99%

Evaluation of capillaries with different inner coatings for DNA analysis using dilute polymer solutions by capillary electrophoresis

Eugênio¹,

Carrilho²

2009

J. Braz. Chem. Soc.

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No presente trabalho, três colunas capilares, uma sem recobrimento interno e duas com recobrimentos internos diferentes foram utilizadas na separação de fragmentos de DNA: poli(vinil álcool) (PVA) e poli(dimetilacrilamida) (PDMA) -ambos de recobrimento covalente -foram comparados para a separação de DNA utilizando soluções poliméricas. As separações foram realizadas usando hidroxietilcelulose (HEC) (90-105 kDa) nas concentrações entre 0,00 e 2,00% m/v. Os resultados indicaram que a eficiência de separação foi maior no capilar de PVA do que no de PDMA, em todas as concentrações de HEC testadas. Ainda, uma resolução superior também foi observada com o capilar de PVA, já que com o capilar de PDMA o formato dos picos não se mostrou reprodutível quando corridas subseqüentes foram realizadas. Contrariamente ao relatado na literatura, nenhuma separação foi conseguida com o capilar sem revestimento interno.In this work three capillary columns, one with uncoated inner wall and two with covalentlybound internal coatings -poly(vinyl alcohol) (PVA) and poly(dimethylacrylamide) (PDMA) -both covalently covered -were used to separate DNA fragments and compared to DNA separation using replaceable polymer solutions. The separations were performed using hydroxyethylcellulose (HEC) (90-105 kDa) in concentrations ranging from 0.00 to 2.00% m/v. The results indicated that the separation efficiency was higher in the PVA capillary than in the PDMA in all evaluated concentrations of HEC. In addition, higher resolution was also observed in PVA-coated capillary since in PDMA the shape of the peaks was not reproducible when subsequent runs were performed. Contrary to what has previously been reported in the literature, no reasonable separation was possible in bare fused silica.Keywords: DNA separation, capillary coating, laser-induced fluorescence, PDMA, PVA IntroductionCapillary electrophoresis (CE) is a separation technique widely used for separation of biological macromolecules. It offers great advantages over slab-gel electrophoresis (SGE) -shorter analysis time, smaller quantity of sample, greater efficiency of separation, and easier automation. 1,2 The first attempts to apply CE to DNA separations were carried out in capillaries filled with cross-linked gels, like polyacrylamide. [3][4][5] Later on, some polymeric solutions were used, known as dynamic or physical gels, which are easily prepared and introduced into the capillary, thus enabling its renewal after each analysis and the use of a single capillary for many separations, without any problem with the subsequent separations. 6 DNA migration mechanisms in polymeric matricesThe Ogston and the reptation models were first proposed to explain DNA migration in slab gels, but they have also shown themselves suitable for polymeric solutions. 7 According to the Ogston theory, the DNA fragment keeps a spherical and rigid conformation of radius Rg. This particle moves across a random net formed by the gel fibers. These fibers form pores with average size ξ, which is a function of the po...

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Capillary electrophoresis of DNA in entangled polymer solutions

Cited by 158 publications

References 17 publications

Separation of double-stranded and single-stranded DNA in polymer solutions: I. Mobility and separation mechanism

Separation of double-stranded and single-stranded DNA in polymer solutions: I. Mobility and separation mechanism

Disposable polyester–toner electrophoresis microchips for DNA analysis

Evaluation of capillaries with different inner coatings for DNA analysis using dilute polymer solutions by capillary electrophoresis

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