Abstract: A coated microcapillary column for high performance electrophoresis is disclosed. A preferred microcapillary includes a fused silica capillary column, the inner surface of the column having an interconnected polymeric coating of a polyvinyl alcohol (PVA) based polymer covalently attached to the column wall by Si--O--Si bonds. The resulting coated microcapillary column has good hydrolytic and pH stability and minimizes electroosmotic flow and interactions between sample components and the capillary wall. Also disclosed are a method of forming a polymeric coating layer for any surface by polymerizing the appropriate organic compounds in an organic solvent and a method of forming a column with a hydrophilic polymeric coating by directly converting an attached hydrophobic polymeric coating material to a hydrophilic polymeric coating material.

Abstract: An electrophoresis separation medium is disclosed comprising a matrix of a copolymer composed of a linear hydrophilic polymer segment having a selected substantially uniform segment length and hydrophobic polymer segments carried on each end. Also disclosed is an electrophoresis method employing the separation medium which permits high resolution separation of polynucleotides, especially in DNA sequencing operations.

Abstract: A device for combined bioaffinity assay and electrophoretic separation is provided, which comprises a capillary system having two stages, a first stage in which bioaffinity interactions of analyte molecules and molecular recognition elements are performed, and a second stage, in which electrophoretic separation of the analyte molecules and subsequent detection of the separated species is accomplished. Within the first capillary stage the molecular recognition elements are attached and immobilized to the inside capillary wall, for example, by adsorption or by covalent binding to the capillary material. The method for combined bioaffinity assay and electrophoretic separation comprises flowing an analyte through a capillary system having two stages. In a first capillary stage the analyte molecules are captured by respective molecular recognition elements present in that stage.

Abstract: A capillary electrophoresis separation matrix for single-stranded nucleic acids, along with methods for using and preparing the matrix, are disclosed. The separation matrix provides denaturing conditions and contains hydroxyethyl cellulose (HEC) in combination with urea, and preferably also includes formamide. The separation matrix may be used for DNA sizing and sequencing applications and provides a single-base resolution to approximately 500 base pairs. The separation matrix is inexpensive, easy to prepare, requires no polymerization steps, and is of low enough viscosity to be pumped easily into and out of capillary tubes for electrophoresis. The low viscosity allows for high throughput of samples and reuse of the capillary tubes for numerous separations.

Abstract: The invention provides a side-entry optical excitation geometry for use in a multiplexed capillary electrophoresis system. A charge-injection device is optically coupled to capillaries in the array such that the interior of a capillary is imaged onto only one pixel. In Sanger-type 4-label DNA sequencing reactions, nucleotide identification ("base calling") is improved by using two long-pass filters to split fluorescence emission into two emission channels. A binary poly(ethyleneoxide) matrix is used in the electrophoretic separations.

Abstract: A capillary column for high performance electrophoretic separation and detection of SDS proteins and system for using the same is disclosed. A preferred column includes a capillary, a static or dynamic layer of coating material on the inner surface of the capillary wall, and a hydrophilic polymer network which is UV-transparent in the tube, filling it. The capillary column is particularly useful in any electrophoresis system requiring UV-transparent materials.

Abstract: Methods for profiling oligosaccharides released from glycoproteins and methods for the electrophoretic separation of high mannose, complex and hybrid oligosaccharides are disclosed. The disclosed methods include first providing an analytical sample which includes at least one released oligosaccharide and introducing the sample into an electrophoretic channel containing an electrophoretic separation medium having a pH less than 8 and greater than 2.5.Then applying a sufficiently high electric field causes the released oligosaccharides to electrophoretically migrate and separate within the electrophoresis channel in accordance with their molecular size or charge to mass ratio. Detecting the separated oligosaccharides provides glycoprotein conjugate profiling information. Preferred embodiments include using an electrophoresis separation medium which includes a lithium acetate buffer having pH of about 4.75 and a gel of about 0.4 wt % polyethylene oxide.

Abstract: A gel pump operated by a stepper motor may be used to deliver fresh gel to one or more capillaries through one or more manifolds and valves. By controlling the valves, fresh gel delivered by the pump will replace the old gel in the capillaries for an automated gel replacement system. In a different setting of the valves, the manifolds may be purged of the old gel prior to delivery of fresh gel to the capillaries. The gel delivery system may also be combined with an electrophoresis system so that sealing connection capable of withstanding high pressure adequate for gel injection need not be frequently broken when the gel is to be replaced. Manifold/reflector assembly is advantageously used in the system that facilitates electrical circuit for electrophoresis and for gel replacement.

Abstract: Disclosed is a substrate useful for separating unmodified and modified mononucleotides and oligonucleotides. The substrate includes at least 12% (weight:volume) polymer in at least 5M urea and at least 32% (volume:volume) organic solvent, the organic solvent being a chemically stable liquid at room temperature and having a dielectric constant of at least 20. Also provided is a method of separating unmodified and modified mononucleotides and/or oligonucleotides utilizing this substrate.

Abstract: Coated capillary electrophoresis columns and methods for their use in electrophoretic separations are disclosed. The coated capillary columns include a length of tubing having an interior surface having an interconnected polymeric coating. The interconnected polymeric coating includes a hydrophobic hydrocarbon polymeric functionality covalently bound to the interior surface and polyvinylalcohol interconnected with the hydrophobic polymeric functionality. Exemplary columns are prepared by causing a Si-OH reactive compound, having a hydrophobic polymeric functionality, to react with Si-OH functionalities on the interior surface of capillary columns. Then causing vinyl acetate to polymerize in contact with the hydrophobic polymeric functionality and hydrolyzing the resulting polyvinylacetate forms a coating of interconnecting hydrophilic and hydrophobic polymers.

Abstract: A Capillary Electrophoresis apparatus and method are disclosed which utilize a meltable plug in an end of the capillary tube to selectively pass small ionic contaminants electrophoretically and retain macromolecular analytes against one end of the plug until the plug is melted. When this plug is melted, the analytes and the contaminants pass through unimpeded. This permits separation of the analytes from the contaminants during electrophoretic separation and enhances instrument resolution.

Abstract: The invention provides a side-entry optical excitation geometry for use in a multiplexed capillary electrophoresis system. A charge-injection device is optically coupled to capillaries in the array such that the interior of a capillary is imaged onto only one pixel. In Sanger-type 4-label DNA sequencing reactions, nucleotide identification ("base calling") is improved by using two long-pass filters to split fluorescence emission into two emission channels. A binary poly(ethyleneoxide) matrix is used in the electrophoretic separations.

Abstract: Separation media for electrophoresis, and methods of filling and flushing of electrophoretic devices such as capillaries are described. By preparing submicron to above-micron sized cross-linked gel particles and using gel swelling equilibrium concepts, such devices can be easily filled and flushed. Gel particles can be prepared by inverse suspension, precipitation and suspension polymerization. These particles can be swollen and collapsed by small changes in temperature, pH, and ionic strength of solvent. Other approaches involve the formation of reversible cross-links by use of polyelectrolyte complexes, chelating agents or copolymers of hydrophobic and hydrophilic repeat units. Finally, reversibly solubilized systems may be used to change the viscosity of the media.

Abstract: The invention provides uncharged water-soluble silica-adsorbing polymers for suppressing electroendoosmotic flow and to reduce analyte-wall interactions in capillary electrophoresis. In one aspect of the invention, one or more of such polymers are employed as components of a separation medium for the separation of biomolecules, such as polynucleotides, polysaccharides, proteins, and the like, by capillary electrophoresis. Generally, such polymers are characterized by (i) water solubility over the temperature range between about 20.degree. C. to about 50.degree. C., (ii) concentration in a separation medium in the range between about 0.001% to about 10% (weight/volume), (iii) molecular weight in the range of about 5.times.10.sup.3 to about 1.times.10.sup.6 daltons, and (iv) absence of charged groups in an aqueous medium having pH in the range of about 6 to about 9.

Abstract: A capillary electrophoresis separation matrix for single-stranded nucleic acids, along with methods for using and preparing the matrix, are disclosed. The separation matrix provides denaturing conditions and contains hydroxyethyl cellulose (HEC) in combination with urea, and preferably also includes formamide. The separation matrix may be used for DNA sizing and sequencing applications and provides a single-base resolution to approximately 500 base pairs. The separation matrix is inexpensive, easy to prepare, requires no polymerization steps, and is of low enough viscosity to be pumped easily into and out of capillary tubes for electrophoresis. The low viscosity allows for high throughput of samples and reuse of the capillary tubes for numerous separations.

Abstract: Described is a method of preparing polyacrylamide gel containing capillary tube wherein acrylamide is polymerized from a liquid to a gel within the tube, the improvement comprising the steps of: a) after the polymerization, annealing the gel inside the tube by cycling the gel/tube at least once in a temperature range from 20.degree. C. to a temperature not to exceed about 75.degree. C. and back to ambient; and b) recovering the stress released gel-filled tube. Preferably, the polymerization commences at the middle of the tube and commences toward the extremities of the tube.

Abstract: A capillary electrophoresis method and apparatus for reducing dispersion of sample fractions are disclosed. The capillary tube in which the electrophoresis is performed has been flared, at least at its sample entrance end, to remove sharp corners which contribute to aberrations in the electric field distribution in a radial direction and result in differential migration of molecules depending on their proximity to the sharp corners. The flared tube, in contrast to a conventional tube, provides a more uniform electric field for electrophoresis and reduces undesired dispersion of the samples.

Abstract: Compositions useful as a separation medium in capillary electrophoresis procedure and methods for their use in capillary electrophoresis are described. The compositions include dry linear polyacrylamide which is capable of hydrating to form an aqueous polymeric system capable of filling an electrophoresis capillary column. Also disclosed are methods for using dry polyacrylamide in capillary electrophoresis. The methods include providing a dry polyacrylamide composition, reconstituting the dry polymer to provide a solution of reconstituted polymer, and causing the reconstituted polymer to fill a capillary electrophoresis column.

Abstract: The invention relates to a process for deactivating the inner surfaces of capillaries for capillary zone electrophoresis and capillary gel electrophoresis. The deactivation is effected by coating the inner surface with polar polymers which initially are water-soluble, whereupon the polymers are subsequently thermally fixed by a formation of water-insoluble polymers. The invention further relates to the capillaries thus produced and to the use thereof for the separation of oligonucleotides, peptides and proteins.

Abstract: A manganese or chromium doped positive plate for a lead acid battery, and a method of manufacturing the same is disclosed. Also disclosed is a lead acid battery containing manganese or chromium in the electrolyte. The doped positive plate has increased cycle life, hardness, and resistance to shedding of the active material.

Abstract: The growth of organisms on the surface of a substrate in contact with water is prevented, while simultaneously preventing corrosion of such substrates. This is accomplished by a method wherein technetium metal is imbedded in, or cast, electroplated or sputtered onto the substrate, or is otherwise included in the surface layer of the substrate being used on or under water.

Abstract: A method for the prevention of fouling by marine growth as well as corrosion on underwater and water-floated objects is disclosed. Technetium metal imbedded in, or cast, electroplated or sputtered onto a metallic substrate, or included in the surface layer of the material is used on or under water in varying concentration levels. The growth of marine organisms on the surface of the material so treated is prevented, while simultaneously preventing corrosion of such substrates.