Factors affecting free zone electrophoresis and isoelectric focusing in capillary electrophoresis

Zhu, Mingde; Hansen, D. L.; Burd, Sam; Gannon, Frank

doi:10.1016/s0021-9673(01)84300-4

Cited by 255 publications

(57 citation statements)

References 11 publications

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“…Although in the past, reports have described CE systems that are based on different polymeric separation media in regard to sizing accuracy and resolution (Cohen et al 1988;Chin and Colburn 1989;Zhu 1989;Heiger et al 1990;Bocek and Chrambach 1991;Sudor et al 1991;Grossman 1992), we describe a novel linear polyacrylamide-derived com-mercially available polymer system specifically developed for high-resolution and high-precision fragment analysis on the ABI 310 Genetic Analyzer. An early version of this separation system, GeneScan polymer (GSP), was shown to be useful for identifying mobility abnormalities in double-stranded bent DNA (Wenz 1994).…”

mentioning

confidence: 99%

High-Precision Genotyping by Denaturing Capillary Electrophoresis

Wenz¹,

Robertson²,

Menchen³

et al. 1998

Genome Res.

119

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Genotyping, as applied to linkage mapping, human identification, or mapping of genetic traits, mandates electrophoretic separation systems that enable a user to identify alleles with high precision to obtain a correct genotype. For 2-bp microsatellites or short tandem repeats (STRs), standard deviations of ±0.3 nucleotide are required to ensure with 99.7% probability the identity or dissimilarity of tested alleles. A complete system, consisting of commercially available laser-induced fluorescence capillary electrophoresis (ABI PRISM 310) and performance optimized polymer 4 (POP-4), was evaluated for microsatellite separations. POP-4 is a low viscosity polymer for use in uncoated fused microbore silica capillaries. It separates DNA fragments that differ in size by 1 nucleotide up to 250 nucleotides and that differ in size by 2 nucleotides for fragments up to at least 350 nucleotides in length in about 30 min. The presence of denaturants and, more importantly, operation at 60°C was mandatory for high-precision and high-resolution sizing operation. Reproducible separation performance was achieved in excess of 100 injections per capillary with resulting standard deviations in the range of 0.04 to 0.17 nucleotide. Comparative sizing of known CEPH (Centre d'Etudes du Polymorphisme Humaine) samples performed at 22 independent test sites showed the usefulness of the system for genotyping with standard deviations of 0.24 nucleotide, or better.

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mentioning

confidence: 99%

High-Precision Genotyping by Denaturing Capillary Electrophoresis

Wenz¹,

Robertson²,

Menchen³

et al. 1998

Genome Res.

119

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“…Protein separations involving 0.03% hydroxypropylmethyl cellulose (HPMC) at pH 2.0 showed separation of the phosphorylated and non-phosphorylated as well as the acetylated and non-acetyl variants of Histone HI (86,87 kb when run at low voltages with good success (91,92,66). Lower voltages may be required when larger fragments are separated due to trapping of the DNA fragments in the polymer network (11).…”

Section: (84)mentioning

confidence: 99%

Optimization of separation and detection schemes for DNA with pulsed field slab gel and capillary electrophoresis

McGregor

1993

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This manuscript has been reproduced from the microfilm master. UMI films the text directly from the original or copy submitted. Thus, some thesis and dissertation copies are in typewriter face, while others may be from any type of computer printer.The quality of this reproduction is dependent upon the quality of the copy submitted. Broken or indistinct print, colored or poor quality illustrations and photographs, print bleedthrough, substandard margins, and improper alignment can adversely affect reproduction.In the unlikely event that the author did not send UMI a complete manuscript and there are missing pages, these will be noted. Also, if unauthorized copyright material had to be removed, a note will indicate the deletion.Oversize materials (e.g., maps, drawings, charts) are reproduced by sectioning the original, beginning at the upper left-hand corner and continuing from left to right in equal sections with small overlaps. Each original is also photographed in one exposure and is included in reduced form at the back of the book.

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“…Much work has been applied to capillary electrophoresis (CE) to develop more sophisticated analytical systems for SBPS. For example, Zhu et al 3 adopted an entangled polymer, poly(ethylene glycol) (PEG), as a substite of a crosslinked sieving polymer such as poly(acrylamide) (PAA). It was reported that 7 proteins (14.2 -205 kDa) were separated within 12 min and also successive runs for 50 times were possible.…”

Section: Introductionmentioning

confidence: 99%

Poly(methylmethacrylate) Microchip Electrophoresis of Proteins Using Linear-poly(acrylamide) Solutions as Separation Matrix

et al. 2008

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Poly(methylmethacrylate) (PMMA) microchip electrophoresis of sodium dodecyl sulfate-protein complexes (SDS-PC) using linear-poly(acrylamide) (L-PA) as a separation matrix was investigated. Prior to electrophoresis, channel walls of PMMA were modified with methylcellulose (MC) to prevent adsorption between channel walls and SDS-PC. Size-based protein separation (SBPS) was successfully performed using the MC-coated microchips with Ferguson plot-fittings. The entangled L-PA solution provided high resolution of peaks of SDS-PC when the concentration of L-PA was increased. Some investigations into the separation mechanism, such as the plot of the logarithm of mobility of each SDS-PC versus the logarithm of the molecular weight of the complex exhibiting linear behavior, indicated that the separation mechanism was dependent on mass discrimination, in accordance with Ogston model.

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Factors affecting free zone electrophoresis and isoelectric focusing in capillary electrophoresis

Cited by 255 publications

References 11 publications

High-Precision Genotyping by Denaturing Capillary Electrophoresis

High-Precision Genotyping by Denaturing Capillary Electrophoresis

Optimization of separation and detection schemes for DNA with pulsed field slab gel and capillary electrophoresis

Poly(methylmethacrylate) Microchip Electrophoresis of Proteins Using Linear-poly(acrylamide) Solutions as Separation Matrix

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