Pluronic copolymer liquid crystals: unique, replaceable media for capillary gel electrophoresis

Rill, Randolph L.; Liu, Yingjie; Winkle, David H. Van; Locke, Bruce R.

doi:10.1016/s0021-9673(98)00522-6

Cited by 81 publications

(60 citation statements)

References 33 publications

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“…17%), the polymers self organize into a transparent liquid crystalline gel of spherical micelles that fill space by packing on a locally cubic lattice [12,13]. Electrophoretic separations of a variety of biological molecules have been reported using Pluronic copolymers as the gel media [13][14][15][16][17][18].…”

Section: Pluronic-f127mentioning

confidence: 99%

A versatile microfabricated platform for electrophoresis of double‐ and single‐stranded DNA

Ugaz¹,

Lin²,

Srivastava³

et al. 2003

Electrophoresis

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We demonstrate a versatile microfabricated electrophoresis platform, incorporating arrays of integrated on-chip electrodes, heaters, and temperature sensors. This design allows a range of different sieving gels to be used within the same device to perform separations involving both single- and double-stranded DNA over distances on the order of 1 cm. We use this device to compare linear and cross-linked polyacrylamide, agarose, and thermo-reversible Pluronic-F127 gels on the basis of gel casting ease, reusability, and overall separation performance using a 100 base pair double-stranded DNA ladder as a standard sample. While cross-linked polyacrylamide matrices provide consistently high-quality separations in our system over a wide range of DNA fragment sizes, Pluronic gels also offer compelling advantages in terms of the ability to remove and reload the gel. Agarose gels offer good separation performance, however, additional care must be exercised to ensure consistent gel properties as a consequence of the need for elevated gel loading temperatures. We also demonstrate the use of denaturing cross-linked polyacrylamide gels at concentrations up to 19% to separate single-stranded DNA fragments ranging in size from 18 to 400 bases in length. Primers differing by 4 bases at a read length of 30 bases can be separated with a resolution of 0.9-1.0 in under 20 min. This level of performance is sufficient to conduct a variety of genotyping assays including the rapid detection of single nucleotide polymorphisms (SNPs) in a microfabricated platform. The ability to use a single microelectrophoresis system to satisfy a wide range of separation applications offers molecular biologists an unprecedented level of flexibility in a portable and inexpensive format.

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Section: Pluronic-f127mentioning

confidence: 99%

A versatile microfabricated platform for electrophoresis of double‐ and single‐stranded DNA

Ugaz¹,

Lin²,

Srivastava³

et al. 2003

Electrophoresis

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“…1d) solutions with an M w of 13 kDa, and with slightly different structures than Pluronic polyol F127, have also been tested extensively as DNA separation matrices [40][41][42][43]. A schematic illustration of the Pluronic F127 micellar structure is shown in Fig.…”

Section: Block Copolymers (Pluronics )mentioning

confidence: 99%

Microchannel DNA sequencing matrices with switchable viscosities

2002

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We review the variety of thermo-responsive and shear-responsive polymer solutions with "switchable" viscosities that have been proposed for application as DNA sequencing matrices for capillary and microfluidic chip electrophoresis. Generally, highly entangled polymer solutions of high-molar mass polymers are necessary for the attainment of long DNA sequencing read lengths (> 500 bases) with short analysis times (< 3 h). However, these entangled polymer matrices create practical difficulties for microchannel electrophoresis with their extremely high viscosities, necessitating high-pressure loading into capillaries or chips. Shear-responsive (shear-thinning) polymer matrices exhibit a rapid drop in viscosity as the applied shear force is increased, but still require a high initial pressure to initiate flow of the solution into a microchannel. Polymer matrices designed to have thermo-responsive properties display either a lowered (thermo-thinning) or raised (thermo-thickening) viscosity as the temperature of the solution is elevated. These properties are generally designed into the polymers by the incorporation of moderately hydrophobic groups in some part of the polymer structure, which either phase-separate or hydrophobically aggregate at higher temperatures. In their low-viscosity states, these matrices that allow rapid loading of capillary or chip microchannels under low applied pressure. The primary goal of work in this area is to design polymer matrices that exhibit this responsive behavior and hence easy microchannel loading, without a reduction in DNA separation performance compared to conventional matrices. While good progress has been made, thermo-responsive matrices have yet to offer sequencing performance as good as nonthermo-responsive networks. The challenge remains to accomplish this goal through the innovative design of novel polymer structures.

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“…Twenty-five percent w/v F127/ No data available addressing toxicity and mutagenicity; should be treated as potential mutagenic/carcinogenic 16TBE solution was filled into the capillary under hand pressure applied by a gas-tight syringe in the cold room (47C) [10]. The polymer solution filled capillary was then equilibrated to room temperature for a few minutes before use.…”

Section: Dye Introduction and Concentration Effectmentioning

confidence: 99%

“…DNA separations can be easily achieved by microchip electrophoresis combined with hydrophilic polymer media. Although the separation of oligos by capillary gel electrophoresis was proven to be superior than that by slab gel electrophoresis and by HPLC [8], the analysis still required a relatively long time period [9,10]. The development of high-speed and high-resolution CE has come to the agenda.…”

mentioning

confidence: 99%

“…Unfortunately, most sieving media at high concentrations have high viscosities, making the filling of polymer solutions into narrow bore capillary tubings very hard or impractical, especially for automated instrumentation. The amphiphilic triblock copolymer Pluronic F127, which forms a cubic macrolattice structure in aqueous solution, is a promising candidate for the separation of small DNA fragments [9,10]. At low temperatures, the poly(propylene oxide) block is hydrophilic and the system acts as a free flowing solution which can be easily injected into the capillary or microchips.…”

mentioning

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Fast separation of single-stranded oligonucleotides by capillary electrophoresis using OliGreen® as fluorescence inducing agent

Zhang

Liang

et al. 2005

Electrophoresis

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The fast separation of oligonucleotide (oligos) sizing marker by CE using OliGreen and including effects due to the concentration of separation medium and urea denaturant is presented. OliGreen dye is found to be more sensitive than ethidium bromide (by a factor of about 6 based on S/N considerations) for the oligos' separations. Higher concentration of F127 in 1xTris-boricacid-EDTA (TBE) up to 30% w/v leads to better resolution of oligos separations. The addition of urea into the separation medium decreases the sensitivity. With an optimized running condition, the oligos sizing marker could be successfully separated with 1-base resolution within 1.3 min by using 30% w/v F127/1xTBE solution as the separation medium at an applied electric field of 800 V/cm in a 3 cm long capillary, the fastest capillary gel electrophoresis separation with high resolution reported to date for oligos in the similar size range.

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Pluronic copolymer liquid crystals: unique, replaceable media for capillary gel electrophoresis

Cited by 81 publications

References 33 publications

A versatile microfabricated platform for electrophoresis of double‐ and single‐stranded DNA

A versatile microfabricated platform for electrophoresis of double‐ and single‐stranded DNA

Microchannel DNA sequencing matrices with switchable viscosities

Fast separation of single-stranded oligonucleotides by capillary electrophoresis using OliGreen® as fluorescence inducing agent

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