Capillary gel electrophoresis

Dolnı́k, Vladislav

doi:10.1002/mcs.1220060402

Cited by 19 publications

(11 citation statements)

References 134 publications

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“…The use of crosslinked gels as separation media in capillaries was largely abandoned by 1993, because of numerous problems including bubble formation, channel clogging, and the inability to load a fresh separation matrix following gel degradation [42]. However, it was observed that highly entangled linear polymer solutions, above C E , behave in a similar manner to crosslinked polymer gels, which provided an alternative that could be readily loaded and removed from capillaries and microfluidic chips [43].…”

Section: Electrophoretic Dna Migration Mechanismsmentioning

confidence: 99%

DNA migration mechanism analyses for applications in capillary and microchip electrophoresis

et al. 2009

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In 2009, electrophoretically driven DNA separations in slab gels and capillaries have the sepia tones of an old-fashioned technology in the eyes of many, even while they remain ubiquitously used, fill a unique niche, and arguably have yet to reach their full potential. For comic relief, what is old becomes new again: agarose slab gel separations are used to prepare DNA samples for "next-gen" sequencing platforms (e.g., the Illumina and 454 machines)-dsDNA molecules within a certain size range are "cut out" of a gel and recovered for subsequent "massively parallel" pyrosequencing. In this review, we give a Barron lab perspective on how our comprehension of DNA migration mechanisms in electrophoresis has evolved, since the first reports of DNA separations by CE (∼1989) until now, 20 years later. Fused silica capillaries, and borosilicate glass and plastic microchips, quietly offer increasing capacities for fast (and even "ultra-fast"), efficient DNA separations. While the channel-by-channel scaling of both old and new electrophoresis platforms provides key flexibility, it requires each unique DNA sample to be prepared in its own micro-or nanovolume. This Achille's heel of electrophoresis technologies left an opening through which pooled-sample, next-gen DNA sequencing technologies rushed. We shall see, over time, whether sharpening understanding of transitions in DNA migration modes in crosslinked gels, nanogel solutions, and uncrosslinked polymer solutions will allow electrophoretic DNA analysis technologies to flower again. Microchannel electrophoresis, after a quiet period of metamorphosis, may emerge sleeker and more powerful, to claim its own important niche applications.

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Section: Electrophoretic Dna Migration Mechanismsmentioning

confidence: 99%

DNA migration mechanism analyses for applications in capillary and microchip electrophoresis

et al. 2009

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“…Capillary Electrophoresis (CE) is also an attractive separation technique without stationary phase, which was originally mainly developed for biomacromolecules such as DNA [18], proteins [19] and polysaccharides [20]. More recently, CE has been used for the characterization of synthetic polyelectrolytes [21][22][23][24][25][26], owing to its rapidity of analysis, minor sample consumption and orthogonal separation mechanism compared to chromatography.…”

Section: Introductionmentioning

confidence: 99%

“…The reptation model [36,37] depicts a snake-like motion of a solute that is too large to pass freely through the pores of the network and must deform / unfold to pass through the matrix. The biased reptation migration (BRM) [21,38,39] describes the forced reptation of a polyelectrolyte under electric field, with orientation and stretching of the chain, leading to electric field-and size-dependent effective electrophoretic mobility, µ [21,39]:…”

Section: Introductionmentioning

confidence: 99%

Characterization of ultrahigh molar mass polyelectrolytes by capillary electrophoresis

Leclercq

Guillard³

et al. 2020

Journal of Chromatography A

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High to ultrahigh molar mass (above 1 million g/mol) anionic poly(acrylic acid-coacrylamide)s are widely used industrial polymers for water treatment and oil drilling. Their properties are strongly related to their charge density and molar mass distributions. However, due to inherent separation limits of SEC with currently available columns (< 5 ×10 6 g/mol) and possible occurrence of chain breakage, and/or adsorption leading to abnormal elution, characterization of unusually high molar masses polyelectrolytes is challenging. In this work, we investigate the use of polymer sieving capillary electrophoresis for the size-based characterization of these high to ultrahigh molar mass polyelectrolytes. By optimizing the operating conditions (electric field, ionic strength, injected polyelectrolyte concentration, nature of the polymer sieving), it has been possible to considerably reduce polyelectrolyte aggregation and to get sufficient size-based selectivity, allowing to obtain the size distribution of the polyelectrolytes over a large range of molar mass from 10 5 up to ~10×10 6 g/mol. The data processing of the raw electropherograms is a key step in the analytical protocol leading to the molar mass distribution. The polyelectrolyte effective mobility in sieving conditions has to be normalized to its free-draining electrophoretic mobility in free solution conditions to take into account possible variability in the charge density between the different samples.

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“…This combined with the impossibility of their replacement resulted in short lifetime of columns regardless of problems with control of the reproducible polymerization inside the capillary. These and some other difficulties were overcome using replaceable entangled polymers 8 that are not covalently crosslinked but form gel networks by physical entanglement of their chains. For this reason, they are often referred to as physical gels.…”

Section: Introductionmentioning

confidence: 99%

Pluronic F‐127 as the buffer additive in capillary entangled polymer electrophoresis: Some fundamental aspects

Křížek

Coufal

Tesařová

et al. 2010

J of Separation Science

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Polymeric macromolecules of well-designed structures and specific properties open promising directions in the capillary entangled polymer electrophoresis. Pluronic F-127, as a thermoassociating polymer, possesses some unique properties that can be utilized in capillary entangled polymer electrophoresis of amino acids, peptides and proteins. In this study, we studied properties of Pluronic F-127 polymer as an additive to BGE for the separation of peptides and proteins. The influence of the thermoassociation on separation selectivity was studied. The addition of Pluronic caused severe instabilities of the electrical current and the signal of the UV detector. This study reveals remarkable positive effect of a low pressure applied to the inlet buffer vial during the analysis, which apparently stabilizes the electrical current and the detector signal. The effect of hydrodynamic flow induced by the pressure applied on the separation efficiency was studied and the significance of this effect was discussed. Pluronic F-127, as a representative of synthetic macromolecules, was compared with dextran, as a representative of natural polymers, in terms of separation power, selectivity and repeatability of migration times.

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Capillary gel electrophoresis

Cited by 19 publications

References 134 publications

DNA migration mechanism analyses for applications in capillary and microchip electrophoresis

DNA migration mechanism analyses for applications in capillary and microchip electrophoresis

Characterization of ultrahigh molar mass polyelectrolytes by capillary electrophoresis

Pluronic F‐127 as the buffer additive in capillary entangled polymer electrophoresis: Some fundamental aspects

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