The Reality of Capillary Electrophoresis

Eby, Michael J.

doi:10.1038/nbt0989-903

Cited by 9 publications

(3 citation statements)

References 7 publications

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“…Capillary electrophoresis (CE) is another technique offering the benefits of rapid separation and integration (4)(5)(6). Since its mechanism of separation is electrophoretic, it is not restricted by limitations associated with analyte-packing interactions, as is HPLC.…”

Section: Introductionmentioning

confidence: 99%

Separation of DNA restriction fragments by capillary electrophoresis using coated fused silica capillaries

Strege¹,

Lagu²

1991

Anal. Chem.

160

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The abilities of several different capillary electrophoresis techniques to separate DNA restriction fragments up to 23,000 bp were investigated. Methods employing electroosmotic flow in an untreated silica capillary were found to provide, at best, only partial resolution of the 23 fragments in a 1-kbp DNA ladder. By coating the inner walls of a silica capillary with poly(acrylamide) and filling these capillaries with buffers containing methylcellulose as a sleving medium, all fragments in the 1-kbp DNA ladder were separated. In addition, this technique facilitated the separation of the very large fragments in a lambda DNA-HindIII digest. Optimum resolution was obtained at low separation potentials using buffers containing at least 0.5% methylcellulose. The performance of this technique, i.e., resolution and quantitation, make capillary electrophoresis a powerful complement to slab gel electrophoresis and may make it a preferred alternative to both agarose gel electrophoresis and HPLC for applications such as the confirmation of plasmid integrity.

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Section: Introductionmentioning

confidence: 99%

Separation of DNA restriction fragments by capillary electrophoresis using coated fused silica capillaries

Strege¹,

Lagu²

1991

Anal. Chem.

160

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“…This provides significant automation advantages over traditional slab gel electrophoresis. CE was first introduced in 1988 [4], but has only recently gained the widespread acceptance in the biopharmaceutical industry that was first anticipated. The amount of sample material and chemical reagents required are extremely low, and the sample amount requirement is approximately 1000-fold less than that required for slab gel methods.…”

Section: Charge Separations By Electrophoresismentioning

confidence: 99%

Process analytics for purification of monoclonal antibodies

Flatman

Alam

Gerard

et al. 2007

Journal of Chromatography B

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“…Additionally, the use of capillaries has promoted electrophoresis into an Abbreviations: TBE,Tris-borate EDTA buffer; UDG, uracil DNAglycosylase instrumental technique. Capillary electrophoresis is now a true counterpart to high performance liquid chromatography and gas chromatography, the more traditional automated analytical methods [4]. …”

mentioning

confidence: 99%

Stability of capillary gels for automated sequencing of DNA

et al. 1992

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Recent interest in capillary gel electrophoresis has been fueled by the Human Genome Project and other large-scale sequencing projects. Advances in gel polymerization techniques and detector design have enabled sequencing of DNA directly in capillaries. Efforts to exploit this technology have been hampered by problems with the reproducibility and stability of gels. Gel instability manifests itself during electrophoresis as a decrease in the current passing through the capillary under a constant voltage. Upon subsequent microscopic examination, bubbles are often visible at or near the injection (cathodic) end of the capillary gel. Gels have been prepared with the polyacrylamide matrix covalently attached to the silica walls of the capillary. These gels, although more stable, still suffer from problems with bubbles. The use of actual DNA sequencing samples also adversely affects gel stability. We examined the mechanisms underlying these disruptive processes by employing polyacrylamide gel-filled capillaries in which the gel was not attached to the capillary wall. Three sources of gel instability were identified. Bubbles occurring in the absence of sample introduction were attributed to electroosmotic force; replacing the denaturant urea with formamide was shown to reduce the frequency of these bubbles. The slow, steady decline in current through capillary sequencing gels interferes with the ability to detect other gel problems. This phenomenon was shown to be a result of ionic depletion at the gel-liquid interface. The decline was ameliorated by adding denaturant and acrylamide monomers to the buffer reservoirs. Sample-induced problems were shown to be due to the presence of template DNA; elimination of the template allowed sample loading to occur without complications.(ABSTRACT TRUNCATED AT 250 WORDS)

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The Reality of Capillary Electrophoresis

Cited by 9 publications

References 7 publications

Separation of DNA restriction fragments by capillary electrophoresis using coated fused silica capillaries

Separation of DNA restriction fragments by capillary electrophoresis using coated fused silica capillaries

Process analytics for purification of monoclonal antibodies

Stability of capillary gels for automated sequencing of DNA

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