Abstract: A solid support and method for immobilization and sequence analysis of polypeptides. In one aspect, the invention is directed to a method for immobilizing a polypeptide on a solid support. In the method, a solid support having surface-attached carboxylic acid groups is reacted with an isoxazolium salt to form an activated support. After the activated support has been washed to remove residual isoxazolium salt and base, the support is dried. The dried support is then contacted with a polypeptide under conditions effective to bind the polypeptide covalently to the support. The immobilized polypeptide can be conveniently sequenced by N- and C-terminal sequencing methods.

Abstract: An apparatus is disclosed for providing capillary electrophoresis, which includes an electronically controlled valve system for automatically introducing a sample into the capillary by means of a vacuum at the end of the capillary tube. This approach of sucking in the sample is extremely accurate and reproducible, and results in a minimum of band broadening. Furthermore, it enables the entire capillary electrophoresis sytem to be easily automated. An automated temperature control system is provided which enables the temperature of the capillary tube (and hence the solvent/solute system) to be controlled during electrophoresis, thereby very directly controlling pH and electrophoretic mobility. In another embodiment, the capillary is prewashed and equilibrated to achieve substantially zero charge on the capillary wall, thereby essentially eliminating electroosmotic flow and substantially improving resolution.

Abstract: Method and kits are provided for detecting one or more target nucleic acids. A first oligonucleotide having a 3' amino group and a second oligonucleotide having a 5' phosphate group are annealed to a contiguous complementary region of a target nucleic acid. Whenever the 3' terminal nucleotides of the first oligonucleotide and the 5' nucleotides of the second oligonucleotide are complementary to the opposing nucleotides on the target nucleic acid, a nucleic acid ligase ligates the first and second oligonucleotides via the formation of a phosphoramidate linkage. The presence of the target nucleic acid is determined by detection of the ligated first and second oligonucleotides.

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: An apparatus to provide a laminar curtain of dry gas over a cryogenic working surface includes a housing duct extending longitudinally from the working surface. A honeycomb member fills a section of the duct adjacent to the working surface, each honeycomb cell having a through channel in the longitudinal direction. A porous foam with through porosity fills a section of the duct adjacent to the honeycomb member oppositely from the working surface. A plenum is adjacent to the porous member oppositely from the honeycomb. Dry gas is supplied to the plenum to flow through the foam and honeycomb to effect the laminar curtain.

Abstract: Method and composition for detecting one or more selected polynucleotide regions in a target polynucleotide. In the method, a mixture of sequence-specific probes are reacted with the target polynucleotide under hybridization conditions, and the hybridized probes are treated to selectively modify those probes which are bound to the target polynucleotide in a base-specific manner. The resulting labeled probes include a polymer chain which imparts to each different-sequence probe, a distinctive ratio of charge/translational frictional drag, and a detectable label. The labeled probes are fractionated by electrophoresis in a non-sieving matrix, and the presence of one or more selected sequences in the target polynucleotide are detected according to the observed electrophoretic migration rates of the labeled probes in a non-sieiving medium.

Abstract: Disclosed is a method for increasing the electro-osmotic flow rate available for a silica surface. In the method, there is provided an electrophoretic channel which is defined by one or more silica surfaces. The surface(s) are contacted with an alkaline aqueous solution containing a solubilized silicate-monovalent metal complex in an amount effective to increase the acidity of the silica surface(s), as evidenced by a reduction in the average bulk pKa of the surface(s). The achieved increase in acidity is greater than would be obtained using an otherwise identical solution lacking said silicate. In one preferred embodiment, the monovalent metal used in the solution is Li.sup.+, Na.sup.+, or K.sup.+. Also disclosed is a method for increasing the acidity of a silica surface, by contacting the surface with an alkaline aqueous solution of the type noted above.

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 spectrometer includes a pair of crossed reflective gratings to effect a spectrally dispersed beam that is focussed to an array detector. The second grating is a plural grating with a surface formed of a first portion and a second portion. The first portion has a groove density for effecting ultraviolet in the dispersed beam. The second portion has a groove density for effecting visible radiation in the dispersed beam. A shutter blocks or exposes the second portion of the grating surface so as to select the first spectral range or the second spectral range for detection. The plural grating surface preferably may be contoured to compensate for aberrations in focussing of the beam to the detector. A computer may be used for selecting spectral lines for analysis, particularly selecting such lines in the second range that are not interfered with by the first range.

Abstract: The compounds are exemplified by the class of sulfoethyl oxygen-substituted carbamates, such compounds being useful as support reagents for automated polynucleotide synthesis of 3'-nitrogen functionalized polynucleotides. The invention includes, in one aspect, a polynucleotide synthesis reagent having the structure: ##STR1## where T is an acid-cleavable hydroxyl protecting group, e.g., 4,4'-dimethoxytritil; Q is a linker connecting the nitrogen and oxygen, e.g., n-hexyl; R.sub.1 is a nitrogen substituent, e.g., hydrogen; R.sub.2 through R.sub.4 are separately hydrogen or lower alkyl; Y is an atom which is electronegative with respect to carbon, e.g., oxygen or sulfur; X.sub.1 is an atom which is electronegative with respect to carbon, e.g., sulfone; Z is a bond or spacer arm, e.g., ethylsuccinate; and W is a derivatized solid synthesis support capable of linking to Z, e.g., an amino-dirivitized controlled pore glass.

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: The invention is directed to a differential analysis method and apparatus wherein a sample and reference are subjected to an externally applied disturbance, such as temperature change, in accord with a prescribed function comprising the sum of a linearly changing part and a periodically changing part, and the measured differential signal is processed into real and imaginary components relating, respectively, to the energy storage and energy loss portions of the signal.

Abstract: A gas chromatographic system includes a column and an injector for delivering into the column a test portion of flow. The injector has an exit passage for discharging a split portion of carrier flow. A flow controller for regulating the inlet flow rate includes a variable outlet restrictor for the split portion, a flow rate detector disposed to detect inlet flow rate, and a feedback flow controller to regulate the outlet restrictor to maintain the inlet flow rate substantially constant. Pressure at the inlet to the injector is regulated by detecting pressure at an outlet and providing feedback control to an inlet restrictor. An integral module for the gas components is used on each side of the injector. Calibration and correction for gas characteristics are provided in the feedback for a chromatographic system and for other flow controller regulation.

Abstract: This invention relates to an improved real-time scanning fluorescence electrophoresis apparatus for the electrophoretic analysis of fluorescently-labeled polynucleotide fragments. The apparatus is characterized by having an electrophoresis chamber containing an electrophoretic separation medium capable of accommodating multiple electrophoresis lanes arranged in a planar array, a fluorescence detector mounted on a translatable stage, a light source for exciting fluorescent molecules, and a computer for collecting data consisting of time, location, fluorescence wavelength and fluorescent intensity information. The improvements herein disclosed include, (i) using a spectral-array detector for detecting the emission light from the fluorescently-labeled polynucleotide fragments including the simultaneous detection of multiple fluorescent labels, and, (ii) a temperature control means to control the temperature of the electrophoretic separation medium during electrophoresis.

Abstract: In a system for measuring variations in thickness of an optical etalon, a light source and a diffraction grating are mounted on a base structure with an axle. A lever arm is affixed to the axle, and a micrometer is held in contact with the lever arm. The grating directs a into an optical path a wavelength of radiation dependent on orientation of the grating. The etalon is supported in the optical path to effect a fringe pattern representing variations in thickness of in the etalon. The orientation is varied with the micrometer so as to vary the wavelength to the etalon and thereby positioning of the fringe pattern across the etalon which is viewed through a microscope. The micrometer measures the variation of orientation and thereby variation in thickness across the interference element.

Abstract: A complete in situ PCR system for amplification of nucleic acids contained in a prepared cell or tissue sample. The containment system for the PCR sample comprises a glass microscope slide, a specimen sample containing the target nucleic acid sequence mounted on the slide, a flexible plastic cover over the sample, and a retaining assembly fastened to the slide and to the cover to retain and seal a reaction mixture against the sample during thermal cycling. The retaining assembly includes a rigid ring on a rim portion of the cover, a cross beam having spaced parallel rails joined by opposite flat ends, and a pair of clips which are pressed over the ends and opposite sides of the slide to fasten the cross beam and cover to the slide.

Abstract: In a spectrophotometer, each of a plurality of source optical fibers is selectively receptive of source radiation and carries the radiation to a corresponding selected liquid sample cell. A corresponding return optical fiber returns transmitted radiation from the sample to a polychromator. For selecting a sample, a switching member holds exposed ends of the optical fibers on a circle coaxial with an axle for rotating to selected positions. Respective optical trains in the instrument direct radiation into and out of the selected pair of fibers. The diameter of a source aperture, the spacing of the aperture from the radiation source, and the source area define a source etendue. The optical fibers have a fiber etendue substantially the same as the source etendue.

Abstract: A method of detecting a target DNA in a sample is provided which includes simultaneously amplifying the target DNA and one or more internal standard DNAs, labeling the target DNA with a first color-producing or color-absorbing label, and labeling each of the one or more internal standard DNAs with a different second color-producing or color-absorbing label, the first and second color-producing and color-absorbing labels being selected so that upon illumination of the sample a first color signal is produced whenever the target DNA is present and a second color signal is produced whenever the target DNA is absent.

Abstract: A rotary scanning apparatus for scanning a plurality of separation lanes where the separation lanes are disposed in a nonplanar array. The scanning apparatus includes an optical detection system for detecting radiation emanating from the separation lanes, the optical detection system including collection optics for collecting and focusing the radiation, a detector for measuring the intensity of the radiation, and a rotary scanner for providing relative movement between the collection optics and the separation lanes.

Abstract: A kit and method for reduction of primer-independent background signals in a DNA replication/detection assay, particularly in a direct in situ PCR amplification/detection assay, for a target DNA sequence in DNA sample, comprising pretreating the DNA sample, containing the target DNA sequence to be replicated/amplified and assayed, with DNA polymerase and one or more dideoxynucleoside triphosphates prior to conducting the replication/amplification steps.