Beckman Instruments, Inc.

Beckman Coulter, Inc.

The United States of America, As Represented by the Secretary, Department of Health and Human Services

Jang B Rampal

Peter J Coassin

Robert S Matson

Edwin M Southern

Charles T Caskey

Serge L Beaucage

Kenneth D Konrad

Edward G Hanna

Ronald M Cook

Stephen W Wunderly

Sushma Rampal

James C Osborne

Steven H Fischer

Carlton B Mc Kinney

Glenn C Sasaki

Gary A Fahle

Paramesewara M Reddy

Patricia S Conville

Jon F Harbaugh

A method for species-specific identification of aspecies is provided. Also disclosed is a method for identifying the species ofpresent in a sample. A device is disclosed for the detection of-specific oligonucleotides are also disclosed, as are kits.

[object Object]

An assay article for detection biopolymers contained in a sample is described. The assay article includes a substrate and a biopolymer directly adsorbed on the surface of the substrate. A plurality of biopolymers may be adsorbed on the surface of the substrate to form an array. Also disclosed is a method of making the assay article. In the preferred method, an aminated polypropylene substrate is used. A biopolymer is contacted with the aminated substrate under a condition sufficient for direct adsorption of the biopolymer on the surface of the substrate. A method of detecting a target biopolymer contained in a sample is also disclosed. In this method, a substrate is contacted with either a probe or target biopolymer under a condition sufficient for a direct adsorption of either the probe or target biopolymer on the substrate to form a probe assay article or a target assay article. Then, the probe assay article is contacted with the target biopolymer, or the target assay article is contacted with the probe biopolymer under a condition that allows the formation of a probe-target complex. Finally, the complex is detected and the presence of the complex is used as a measurement for the presence or the amount of the biopolymer target contained in the sample.

Immobilization of biopolymers to aminated substrates by direct adsorption

Disclosed herein is a method for attaching pre-synthesized oligonucleotides to a polypropylene support medium. Most preferably, a polypropylene film is aminated by a plasma discharge in the presence of ammonia gas. An oligonucleotide having a terminal phosphate is activated in the presence of an imidazole and a carbodiimide to form a phosphorimidazolide. The activated oligonucleotide becomes immobilized by forming a phosphoramidate bond with the aminated polypropylene. The invention can be used to construct oligonucleotide arrays for hybridization assays.

Covalent attachment of biomolecules to derivatized polypropylene supports

An improved method for detecting hybridized biomolecules is provided which comprises the steps of, providing a plurality of probe biomolecules on a solid support surface; pre-treating the plurality of probe biomolecules with an effective amount of a pre-treatment solution to produce a signal having an increased intensity of at least two (2) times, as compared to an identical detection method that does not include the pre-treating step; applying a buffer solution to the probe biomolecules; hybridizing the probe biomolecules with a plurality of target biomolecules to form a hybridized complex; and, developing and detecting the signal intensity. The developing step of the method may further include the steps of, treating the hybridized complex with a conjugating solution, and treating the conjugated, hybridized complex with a detection reagent.

Hybridization detection by pretreating bound single-stranded probes

A solid support based hybridization assay is provided which allows for the systematic and reproducible analysis of repeat and tandem repeat oligonucleotide sequences of DNA and RNA by hybridization to a reverse dot blot array comprising strings of such repeats complementary to those found in particular nucleic acid targets (e.g., analyte PCR product). An addressable library (i.e., an indexed set) of complementary repeats is synthesized on a suitable support. Preferably, the support comprises a low fluorescent background support, thereby facilitating the use of non-radioisotopic modes of detection (such as fluorescence or chemiluminescence); particularly suitable in this regard is an aminated polypropylene support or similar material. Preferred arrays permit screening of DNA and RNA samples for complete sets of particular types of nucleotide repeat sequences (e.g., all nucleotide doublet or triplet repeats).

Oligonucleotide repeat arrays

Disclosed herein are surface activated, organic polymers useful for biopolymer synthesis. Most preferably, aminated polypropylene is used for the synthesis of oligonucleotides thereto, and these devices are most preferably utilized for genetic analysis of patient samples.

Surface activated organic polymers useful for location - specific

Disclosed herein are surface activated, organic polymers useful for biopolymer synthesis. Most preferably, aminated biaxially oriented polypropylene is used for the synthesis of oligonucleotides thereto, and these devices are most preferably utilized for genetic analysis of patient samples.

Biopolymer synthesis using surface activated biaxially oriented

A sample having both positively and negatively charged components may be separated during the same run by injecting a plug of sample at both ends of the capillary and applying an electric field in the capillary, where there is insufficient electroosmotic flow in the capillary to reverse sample migration direction, such as in the case of a gel-filled column.

Capillary electrophoretic system for separation of samples containing

Inverse Linkage Oligonucleotides ('ILO') useful in enzymatic process are disclosed. Particularly preferred ILOs are amenable to enzymatic elongation from either, or most preferably, both termini. In a particularly preferred embodiment, each terminus of an ILO has an enzymatically functional 3' group. Accordingly, under appropriate conditions and in the presence of, e.g., dNTPs, enzyme, sample DNA, and ILO comprising a first region complementary to a first region of the sample DNA and a second region complementary to a second, different region of the sample DNA, exponential amplification of the sample DNA can be effectuated.

Inverse linkage oligonucleotides for chemical and enzymatic processes

Procedures such as oligonucleotide, peptide or protein syntheses or sequencing, or immunological or other biospecific assays involving long sequential series of chemical reactions are performed on a solid phase support retained in a pipette tip. The manipulations involved in bringing the solid support into contact with the various reagents or reactants involved in the procedure are performed and controlled by automated instrumentation.

Solid phase system for sequential reactions

An automated method and apparatus for performing biopolymer synthesis on a two-dimensional support surface whereby a two-dimensional matrix or array of biopolymers are obtained on the surface. An applicator in the form of a thick plate having in a surface at least one cavity in the form of an open chamber or channel is used for applying reagents to the surface of a solid phase support material. The applicator is positioned over the solid support material with the surface having the cavity sealed against the material surface. For each cavity, reagents for synthesis are introduced sequentially into one end of the cavity and collected from the other end of the cavity. A one-dimensional array of biopolymers are thus formed on the support material, where each element of the array contains a population of biopolymers having identical sequence. Further synthesis of additional biopolymers is performed by linearly moving or rotating the applicator relative to the support material and carrying out synthesis procedure so that another one-dimensional matrix or array of biopolymers are formed at an offset position to the previous array. At each overlapping region of the two arrays, a cell of biopolymers are formed which each comprises a strand from the first array and a strand from the second array. The result is a two-dimensional array of biopolymers of different sequences at each discrete cell. The arrays of discrete cell containing different biopolymers may be used to conduct hybridization reaction analysis of a biological sample.

Method and apparatus for creating biopolymer arrays on a solid support

The present invention is directed to a synthesis reaction column which can be conveniently implemented in post synthesis procedures. The reaction column is configured to allow easy connection to a syringe and post synthesis reagent container without requiring connection adapters. In one embodiment, the reaction column configured in the form of a cylindrical column containing the solid-phase support and having a narrow bore tube extending at one end. The tube functions as a piercing tip for accessing a septum sealed vial containing post synthesis reagent. In another embodiment, the reaction column is configured in the form of a cylindrical barrel enclosing the solid-phase support and having a threaded mount at one end. To complement the reaction column for post synthesis procedures, a vial having a complementary threaded mount at the opening is used for containing a reagent. This vial is conveniently threaded onto the end of the barrel without adapters. Consequently, post synthesis procedures can be implemented conveniently. Both embodiments may be conveniently connected to the reagent delivery system in an automated instrument for carrying out synthesis procedures, as well as post synthesis procedures.

Synthesis reaction column

Method for substituting a substitution compound for the hydrogen of a support bound primary hydroxyl group. The support bound primary hydroxyl group is treated in an organic, substantially anhydrous solvent in the presence of a tertiary amine base and the substitution compound. The substitution compound has a positive moiety and a negative moiety. The positive moiety is selected from the group consisting of trityl, pixyl, and derivatives thereof. The reagent is selected from the group consisting of tetraalkylammonium salts, inorganic salts, and mixtures thereof. Each reagent has a cation and an anion moiety. The cation moiety of the reagent is more electropositive than the positive moiety of the substitution compound. This relationship enables the reagent to attract the negative moiety of the substitution compound.

Growing community of inventors

Yorba Linda, CA, United States of America

Jang B Rampal