Abstract: A method is described for retrieval of phylogenetically informative DNA sequences which comprise searching for a highly divergent segment of genomic DNA surrounded by two highly conserved segments, designing the universal primers for PCR amplification of the highly divergent region, and amplifying the genomic DNA by PCR technique using universal primers.
Abstract: Tubing for a brake system, fuel system or hydraulic system in automotive vehicles which includes an inside steel tube having an outer surface, a zinc coating layer on the outer surface, chromated layer overlying the zinc coating layer and a plastic jacket layer overlying the chromated layer wherein the plastic jacket layer is composed of a polyamide on the chromated layer, the polyamide preferably being laurinlactam and the chromated layer preferably being olive-colored. The method of tubing manufacture including the steps of heating a steel tube having an outer surface with the zinc coating and chromate present thereon to an elevated temperature; the elevated temperature being within about .+-.30.degree. C. of the extrusion temperature of polyamide material to be applied to the tube; extruding polyamide material onto the surface of the heated tube; and quenching the tube in a liquid bath after the polyamide has been applied.
Abstract: This method for detecting a target nucleic acid sequence involves amplification and detection in the same vessel and comprises: (a) amplification of the target nucleic sequence in a vessel which is provided with a solid phase capture probe comprising a nucleic acid sequence capable of hybridizing to at least a portion of said amplified target nucleic acid sequence, said capture probe being incapable of participating or not participating in standard nucleic acid sequence amplification processes, (b) bringing a sample suspected of comprising said target nucleic acid sequence into contact with said capture probe under conditions which allow said amplified target nucleic acid sequence to be bound by said capture probe, and (c) detecting the presence of bound target nucleic acid sequence.
Type:
Grant
Filed:
January 24, 1995
Date of Patent:
December 15, 1998
Assignees:
University of Australia, Adelaide Children's Hospital
Inventors:
Raymond John Harris, Charles Phillip Morris
Abstract: Methods are provided for the hybridization of a target polynucleotide with a conjugate. The conjugate comprises an oligonucleotide covalently bound to a linker that is covalently bound to an amino acid or a polyamide. The oligonucleotide portion of the conjugate comprises an anti-sense sequence which is complementary to and hybridizes with the target polynucleotide. The method is useful for detecting the presence of viral or bacterial target polynucleotides.
Type:
Grant
Filed:
October 27, 1997
Date of Patent:
December 8, 1998
Assignee:
Howard Florey Institute of Experimental Physiology and Medicine
Inventors:
Jim Haralambidis, Geoffrey William Tregear
Abstract: The present invention provides an in situ monitoring of the decomposing activity against oligonucleotides target in a biological tissue. A single-chain oligonucleotide target comprises an appropriate number of nucleic acid bases to be examined with an energy donor and an energy acceptor respectively at its 5'- and 3'-terminals. Monitoring of the fluorescence changes of the target after injection into a biological tissue, particularly the fluorescent resonance energy transfer(FRET) phenomena between the energy donor and acceptor, indicates whether the oligonucleotide is not decomposed yet.
Abstract: Methods employing internal oligonucleotide standards in isothermal nucleic acid amplification reactions to determine the efficacy of the amplification reaction and to quantify pre-amplification target levels.
Abstract: Compositions and methods are provided for the modulation of abnormal expression of .beta.-amyloid. Oligonucleotides are provided which are specifically hybridizable with RNA or DNA encoding .beta.-amyloid. Oligonucleotides specifically hybridizable with a translation initiation site, codon 717, codon 670 or codon 671 of .beta.APP are provided. Such oligonucleotides can be used for diagnostics as well as for research purposes. Methods are also disclosed for modulating .beta.-amyloid expression in cells and tissues using the oligonucleotides provided, and for specific modulation of expression of the mutant .beta.APP gene. Methods for diagnosis, detection and treatment of diseases associated with abnormal .beta.APP expression are also disclosed.
Type:
Grant
Filed:
October 28, 1994
Date of Patent:
November 17, 1998
Assignee:
Isis Pharmaceuticals, Inc.
Inventors:
Brett P. Monia, Susan M. Freier, David J. Ecker
Abstract: A region of the Chlamydia trachomatis ltuB gene has been identified which is useful for performing amplification assays to determine specifically whether C. trachomatis is present in the sample being tested. Oligonucleotides useful for performing thermophilic Strand Displacement Assay (tSDA) reactions on this gene are disclosed. The disclosed oligonucleotides can be used in an assay which is specific for all strains of C. trachomatis and which does not show crossreactivity with the genomes of other microorganisms or with human DNA.
Abstract: A method of mutagenesis by which a predetermined amino acid is introduced into each and every position of a selected set of positions in a preselected region (or several different regions) of a protein to produce library of mutants. The method is based on the premise that certain amino acids play crucial role in the structure and fuction of proteins. Libraries can be generated which contain a high proportion of the desired mutants and are of reasonable size for screening. This libraries can be used to study the role of specific amino acids in protein structure and function and to develop new or improved proteins and polypeptides such as enzymes, antibodies, single chain antibodies and catalytic antibodies.
Abstract: Sequencing of a selected region of a target nucleic acid polymer in a genomic DNA sample can be performed in a single vessel by combining the sample with a sequencing mixture containing a primer pair, a thermally stable polymerase such as Thermo Sequenase.TM. which incorporates dideoxynucleotides into an extending nucleic acid polymer at a rate which is no less than about 0.4 times the rate of incorporation of deoxynucleotides, nucleotide feedstocks, and a chain terminating nucleotide. The mixture is processed through multiple thermal cycles for annealing, extension and denaturation to produce a product mixture which is analyzed by electrophoresis.
Type:
Grant
Filed:
July 19, 1996
Date of Patent:
November 3, 1998
Assignee:
Visible Genetics Inc.
Inventors:
James Leushner, May Hui, James M. Dunn, Marina T. Larson
Abstract: A method of analyzing an amplified gene, including determining its copy number, involves subtractive hybridization of two cDNA libraries, one from the tissue of interest and the other containing biotinylated cDNA from normal tissue, where the annealed cDNA is removed by means of magnetic beads coated with streptavidin or avidin. The cDNA isolated after subtractive hybridization represents amplified DNA, and it is analyzed to determine what gene(s) were amplified. Furthermore, the copy number of the gene(s) can be estimated. The copy number thus determined can be correlated to the severity of a pathogenic state, to its prognosis, or to treatment efficacy.
Type:
Grant
Filed:
August 9, 1996
Date of Patent:
October 27, 1998
Assignee:
The United States of America as reprsented by the Department of Health and Human Services
Abstract: An RNA amplification method is particularly useful for diagnosing bacterial or viral infections or genetic disorders and for cell typing. The method includes the steps of denaturing a solution containing RNA, synthesizing a first cDNA strand from a suitable primer in the presence of reverse transcriptase, denaturing the heteroduplex formed, synthesizing a second cDNA strand from a second primer in the presence of DNA polymerase and then subjecting the cDNA formed to a sufficient number of amplification cycles. All the reactants and solvents are first placed in the same container to provide a single manipulation step that avoids the risk of contamination.
Type:
Grant
Filed:
March 31, 1997
Date of Patent:
October 6, 1998
Assignee:
Bio Merieux
Inventors:
Francois Mallet, Guy Oriol, Bernard Mandrand
Abstract: Compounds referred to herein as oligonucleotide clamps are provided that stably bind to target polynucleotides in a sequence-specific manner. The oligonucleotide clamps comprise one or more oligonucleotide moieties capable of specifically binding to a target polynucleotide and one or more pairs of binding moieties covalently linked to the oligonucleotide moieties. In accordance with the invention, upon annealing of the oligonucleotide moieties to the target polynucleotide, the binding moieties of a pair are brought into juxtaposition so that they form a stable covalent or non-covalent linkage or complex. The interaction of the binding moieties of the one or more pairs effectively clamps the specifically annealed oligonucleotide moieties to the target polynucleotide.
Abstract: A labeled nucleic acid probe comprising a single-stranded oligonucleotide probe having a nucleic acid sequence complementary to a specific nucleic acid sequence in a target nucleic acid and an intercalating fluorochrome bound to said probe is used to detect the target nucleic acid by a convenient, single-stage method in a homogeneous system. The formation of a complementary bond between the probe and the target nucleic acid can be detected and the amount of the resulting complementary binding product determined without requiring any extra step such as for removing the excess probe which has not participated in the complementary binding.
Abstract: Sets of fluorescent energy transfer labels and methods for their use in multi component analysis, particularly nucleic acid enzymatic sequencing, are provided. In the subject sets, at least two of the labels are energy transfer labels comprising a common donor and acceptor fluorophore in energy transfer relationship separated by different distances and capable of providing distinguishable fluorescence emission patterns upon excitation at a common wavelength. The subject labels find particular use in a variety of multi-component analysis applications, such as probes in FISH and multi array analyses, as well as primers in nucleic acid enzymatic sequencing applications.
Abstract: A method of characterizing a test sample of genomic DNA which method comprises amplifying a tandemly repeated region, comprising more than one type of repeat unit, as far as internal repeat units of a specific type so as to generate a set of amplification products which identify the relative positions of the internal repeat units within the tandemly repeated region, and separating the set of amplification products to provide a sample code. The sample codes are suitable for computerized storage on, and retrieval from, a database. The invention also provides a novel method for the detection of diagnostic base sequences in one or more nucleic acids contained in a sample.
Abstract: An aqueous composition containing primers for opposing strands of two or more target nucleic acids can be used in polymerase chain reaction to provide simultaneously rapid and efficient amplification and detection of those nucleic acids. The primers for each target DNA differ in length by no more than 5 nucleotides and have a T.sub.m within the range of from about 65.degree. to about 74.degree. C., while the T.sub.m 's are within about 5.degree. C. of each other. Such compositions are useful in diagnostic test kits and methods for amplification and detection of multiple nucleic acids, or in "multiplexing", using multiple capture probes. All of the capture probes have T.sub.m 's which are greater than 50.degree. C. and are within 15.degree. C. of each other.
Type:
Grant
Filed:
June 8, 1995
Date of Patent:
September 22, 1998
Assignee:
Johnson & Johnson Clinical Diagnostics, Inc.
Inventors:
Thomas J. Cummins, Susan Melissa Atwood, Lynn Bergmeyer, John Bruce Findlay, John W. H. Sutherland, JoAnne H. Kerschner