Abstract: The present invention provides methods for detecting amplified or unamplified nucleic acid target sequences at increased temperatures by changes in fluorescence polarization. The decrease in fluorescence polarization associated with hybridization of oligonucleotides at higher, more stringent, temperatures is overcome by including a double-stranded DNA binding protein in the assay. At elevated temperatures, the double-stranded DNA binding protein restores, and often enhances, the magnitude of the change in fluorescence polarization associated with single- to double-stranded conversion of an oligonucleotide probe or primer.

Abstract: Improved homogenous diagnostic assay methods and labels for detecting an analyte in a medium when the analyte is a member of a specific binding pair. The methods and labels provide procedures for reducing background and increasing sensitivity. The binding partner of the analyte is labeled with a substance, the stability of which detectably changes whenever said analyte is bound as a member of the specific binding pair. In a closely related system, the analyte is labeled with a substance susceptible to differential degradation depending on whether or not the analyte is bound as a member of its specific binding pair. After incubation and selective degradation or chemical or biochemical alteration, the amount of analyte bound is detected by measuring either the stability change or the extent of degradation of the label.

Abstract: This invention is directed to methods for determining a nucleotide sequence of a nucleic acid using positional sequencing by hybridization, and to the creation of nucleic acids probes which may be used with these methods. This invention is also directed to diagnostic aids for analyzing the nucleic acid composition and content of biological samples, including samples derived from medical and agricultural sources.

Abstract: The present invention provides a method for simplifying and significantly enhancing the sensitivity of nucleic acid hybridization assays. A method is described whereby a single-stranded primary nucleic acid sequence that includes a region of sequences complementary to a single-stranded target nucleic acid sequence is hybridized to the target molecule. Stability of the double-stranded complex thereby formed can be enhanced by using RNA as the probe if DNA is the target or DNA as the probe if RNA is the target. The probe-target complex is subsequently immunocaptured for detection. After washing away extraneous material, a secondary nucleic acid sequence containing many repeating sequence units is hybridized to the probe component of the immobilized probe-target complex. Detection occurs following hybridization of many labelled nucleic acid sequence probes to each of the repeating sequence units of a nucleic acid amplification probe.

Abstract: The present invention relates to a method of identifying DNA encoding an osteoclast-specific or -related gene product. The method comprises hybridizing DNA with a stromal cell+, osteoclast+probe, or with cDNA or mRNA from an osteoclastoma, and with a stromal cell+, osteoclast-probe, or with cDNA or mRNA from stromal cells; and identifying DNA which hybridizes to the stromal cell+, osteoclast+probe, or to cDNA or mRNA from osteoclastoma, but not to the stromal cell+, osteoclast-probe, or to cDNA or mRNA from stromal cells.

Abstract: This invention relates to the use of functional reporter molecules in the detection and measurement of targets in a sample. The invention is predicated on the utilization of a transcription step between the production of an appropriate reporter molecule and replication based amplification in order to increase the number of detectable species as an indirect reference to target.

Abstract: Oligonucleotides of formula(X).sub.a -(6N-palindrome).sub.n -(Y).sub.b ( 1)wherein:"6N-palindrome" represents TACGTA, ACGTAT, CGTATA, GTATAC, TATACG or ATACGT (read in the conventional notation with the 5'-end at the left);n is an appropriate integer of at least 2;X is a length of appropriate 5'-end or tailing DNA and a is 0 or 1; andY is a length of appropriate 3'-end or heading DNA and b is 0 or 1;or any labelled form thereof or any derivative thereof which retains the ability of the 6N-palindrome sequence to anneal or hybridize to its complementary sequence are useful in assay of encephalopathies such as scrapie, BSE or CJD. Preferably they are used as PCR primers to amplify DNA in the sample. PCR product can be detected by restriction with an appropriate enzyme and comparing the restricted and unrestricted products.

Abstract: Nucleic sequences from the genome of Salmonella typhi include all or part of the genetic information required for the in vitro infection of cultured HeLa cells by Salmonella bacteria. Polypeptides encoded by these nucleic sequences are also described, as is the use of said polypeptides and nucleic sequences for implementing methods of in vitro Salmonella detection in biological samples which are thought to contain it.

Abstract: A new gene from Bacillus cereus has been characterized which codes for a protein conferring resistance to the antibiotic zwittermicin A produced by some strains of the bacteria. The resistance gene, designated zmaR, is found to be present in almost all strains of the bacteria which are known to natively have the ability to synthesize zwittermicin A, and thus allows tests for the presence of the gene to serve as a test to be used with new strains to assay for zwittermicin A production. The gene also has use in increasing production of the antibiotic by producing strains.

Abstract: In a method for molecular-biological analysis of genetic material, a genomic DNA or RNA preparation is tested for the presence of mutations. The processed DNA or RNA preparation is contacted with a first set(s) of interconnected solid phase members (5) supporting oligonucleotide primers to bind a defined DNA or RNA fragment that may contain a stable mutation to each solid phase member, and/or with a second set(s) of interconnected solid phase members (5) supporting oligonucleotide primers to bind a defined DNA or RNA fragment that may contain an unstable mutation or several stable mutations to each solid phase member. The solid phase members (5) of the first set(s) are introduced into a matching first set(s) of interconnected receptacles (7) with reaction mixtures for producing products which contain an incorporated marker when a supported DNA or RNA fragment has a mutation.

Abstract: Stabilized enzyme compositions for use in nucleic acid amplification. Compositions are provided for the stabilization of one or more enzymes in a single stabilized formulation. Additional compositions incorporate a dried, stabilized enzyme mixture together with necessary cofactors and enzyme substrates in a single container for use upon rehydration. Also disclosed are methods for making and using stabilized enzyme compositions and kits for nucleic acid amplification incorporating the disclosed compositions.

Abstract: The invention provides fast and highly accurate mass spectrometer based processes for detecting a particular nucleic acid sequence in a biological sample. Depending on the sequence to be detected, the processes can be used, for example, to diagnose (e.g. prenatally or postnatally) a genetic disease or chromosomal abnormality; a predisposition to a disease or condition (e.g. obesity, artherosclerosis, cancer), or infection by a pathogenic organism (e.g. virus, bacteria, parasite or fungus).

Abstract: The invention provides a method of nucleic acid sequence analysis based on repeated cycles of ligation to and cleavage of probes at the terminus of a target polynucleotide. At each such cycle one or more terminal nucleotides are identified and one or more nucleotides are removed from the end of the target polynucleotide, such that further cycles of ligation and cleavage can take place. At each cycle the target sequence is shortened by one or more nucleotides until the nucleotide sequence of the target polynucleotide is determined. The method obviates electrophoretic separation of similarly sized DNA fragments and eliminates the difficulties associated with the detection and analysis of spatially overlapping bands of DNA fragments in a gel, or like medium. The invention further obviates the need to generate DNA fragments from long single stranded templates with a DNA polymerase.

Abstract: A method for isolating mRNAs as cDNAs employs a polymerase amplification method using at least two oligodeoxynucleotide primers. In one approach, the first primer contains sequence capable of hybridizing to a site immediately upstream of the first A ribonucleotide of the mRNA's polyA tail and the second primer contains arbitrary sequence. In another approach, the first primer contains sequence capable of hybridizing to a site including the mRNA's polyA signal sequence and the second primer contains arbitrary sequence. In another approach, the first primer contains arbitrary sequence and the second primer contains sequence capable of hybridizing to a site including the Kozak sequence. In another approach, the first primer contains a sequence that is substantially complementary to the sequence of a mRNA having a known sequence and the second primer contains arbitrary sequence.

Abstract: A process of identifying and disrupting bacterial DNA sequences that contribute to the evolution of new lytic bacteriophages is described. Vectors and recombinant bacteria for use in producing fermentative starter cultures and culture media resistant to the appearance of new phages, and methods of producing such vectors and recombinant bacteria, are described.

Abstract: Qualitative and quantitative methods for detecting the presence of double-stranded, non-5'-phosphorylated DNA in samples that may also contain 5'-phosphorylated DNA and/or single-stranded DNA are described. These methods involve treating the sample with an enzyme that specifically degrades 5'-phosphorylated DNA together with an enzyme that specifically degrades single-stranded DNA. More specifically, methods are described for the detection of the products of DNA amplification reactions, such as the polymerase chain reaction (PCR), wherein post-amplification enzyme digestion substantially reduces or eliminates background signals that are apparently caused by the presence of template DNA and primers in the sample after amplification is complete.

Abstract: An exonuclease-based method for joining and/or constructing two or more DNA molecules. DNA fragments containing ends complementary to those of a vector or another independent molecule were generated by the polymerase chain reaction. The 3' ends of these molecules as well as the vector DNA were then recessed by exonuclease activity and annealed in an orientation-determined manner via their complementary single-stranded regions. This recombinant DNA may be transformed directly into bacteria without a further ligase-dependent reaction. Using this approach, recombinant DNA molecules are constructed rapidly, efficiently and directionally. This method can effectively replace conventional protocols for PCR cloning, PCR SOEing, DNA subcloning and site-directed mutagenesis.

Abstract: The present invention provides nucleic acids encoding consensus hemagglutinin and consensus fusion polypeptides which contain at least one amino acid substitution relative to the Moraten vaccine strain that is found in at least one strain of wild-type measles virus. Nucleic acid reagents useful for detecting and differentiating wild-type measles strains by polymerase chain reaction and a method utilizing the same are provided. Recombinant vectors comprising nucleic acids encoding the consensus hemagglutinin and/or the consensus fusion polypeptides are also provided.

Abstract: According to the process of the invention, sets of restriction fragments from the DNA of an individual are prepared, these restriction fragments being separated by size. Probes are prepared by enzymatic hydrolysis of DNA from a genome bank of the species to which this individual belongs, separation of the fragments obtained as a function of their size, labeling these probes and placing them in contact, under hybridization conditions, with the aforementioned sets of restriction fragments. Finally, selection is made of the probes (capable of hybridizing with said set of restriction fragments) which do not give hybridization profiles identical to those obtained with known probes recognizing variable number tandem repeat regions. Applications for the probes thus obtained include, particularly, processes for identifying an individual, consanguinity testing, and investigating the origin of a seed.

Abstract: The present invention provides methods of detecting a change in the length of an oligonucleotide labeled with a light-emitting label by measuring the light emission in the presence of a DNA binding compound that interacts with the label to modify the light emission of the label, wherein the degree of interaction depends on the length of the oligonucleotide. The methods are applicable in general to nucleic acid sequence detection assays that utilize a reaction that results in the selective cleavage of hybridized oligonucleotide probes, and, in particular, to amplification/detection assays wherein hybridized probes are cleaved concomitant with primer extension.