Abstract: This invention discloses novel immunoassay termed an ELONA, employing nucleic acid ligands as capture molecules and/or detector molecules.

Abstract: This invention involves a method of altering and regulating the gene expression of cells, by means of contacting the cells with a nuclease capable of degrading extra-cellular DNA and/or RNA. The nuclease will degrade the extra-cellular nucleic acids (NA) into nucleotides or oligonucleotides which are too short to have substantial affinity for the chromosomal DNA.By means of this method, cultures of cells have been created with desirable properties, including greater phenotypic uniformity and higher levels of cell reproduction.This invention also comprises a culture of cells which has been treated by this method, and cells descended from cells which have been treated according to this method.

Abstract: A method for analyzing a sample oligonucleotide sequence is described. The method comprises contacting the sample oligonucleotide sequence with an anchor sequence which comprises an immobilized oligonucleotide sequence which hybridizes with the sample. The sample is also contacted with a probe comprising an oligonucleotide sequence which hybridizes to a target oligonucleotide sequence to be detected in a suitable buffer to form a complex. The complex is subjected to a field which moves unbound oligonucleotide sequences away from the anchor sequence in the direction of the field, and preferably, extends the sample sequence. Whether the probe is bound to the sample oligonucleotide sequence, and preferably, the position of the probe, is determined to determine whether the target oligonucleotide sequence is present in the sample. The method can be used for mapping, for identity typing, and to determine whether a test oligonucleotide sequence is present in the sample.

Abstract: Methods are described for the identification and preparation of DNA ligands to the HIV-1 reverse transcriptase protein. Included in the invention are specific ssDNA ligands to HIV-1 reverse transcriptase identified by the SELEX method. Also included are ssDNA ligands that inhibit HIV-1 reverse transcriptase.

Abstract: Methods for inhibiting the secretion of .beta.-amyloid peptide (.beta.AP) from cells comprise administering to the cells certain compounds which inhibit the activity of an approximately 31 kD protease involved in .beta.AP secretion. The 31 kD protease has been designated Cathepsin Y. Screening methods for .beta.AP inhibitors rely on determining the activity of test compounds in the presence of Cathepsin Y and a suitable peptide substrate. This invention is also directed to a nucleic acid sequence that encodes Cathepsin Y and the expression and isolation of Cathepsin Y.

Abstract: A method of detecting and identifying specific human nucleic acid sequences contained in human nucleic acids in a human blood or tissue sample, comprising: (1) amplifying at least one portion of the specific nucleic acid sequence present in the sample; (2) transcribing the amplification product with an RNA polymerase to produce multiple RNA copies of each copy of specific nucleic acid sequence comprising the amplification product; and (3) identifying the transcription product.

Abstract: A method of preparing a homogeneous alkaline phosphatase-oligonucleotide probe conjugate having high specific enzyme activity for use in nucleic acid hybridization assays is disclosed. Indirect, competitive nucleic acid hybridization assay formats are also described.

Abstract: Methods are provided for substantially reducing background signals encountered in nucleic acid hybridization assays. The method is premised on the elimination or significant reduction of the phenomenon of nonspecific hybridization, so as to provide a detectable signal which is produced only in the presence the target polynucleotide of interest. In addition, a novel method for the chemical synthesis of isoguanosine or 2'-deoxy-isoguanosine is provided. The invention also has applications in antisense and aptamer therapeutics and drug discovery.

Abstract: A method for breeding mutagenized mice that permits detection of genetic loci that can modify a known index phenotype involves crossing a mutagenized founder strain and a second strain of mice carrying an allele at a locus that confers the index phenotype. In the test generation, clusters of individuals are observed to deviate from the typical phenotype. The genetic material and molecules encoded thereby can be obtained using available methods.

Abstract: Assays for determining cartilage synthesis associated with osteoarthritis are presented. The assays are based on detection of type IIA procollagen or propeptide, or type IIA mRNA in a tissue or fluid sample from a non-neonatal individual being tested. Since type IIA procollagen is not found in normal non-neonatal individuals, type IIA procollagen and the mRNA which encodes it are unique markers for osteoarthritis which exhibits neonatal like cartilage synthesis as part of the disease syndrome.

Abstract: An improved nucleic acid hybridization process is provided which employs a modified oligonucleotide and improves the ability to discriminate a control nucleic acid target from a variant nucleic acid target containing a sequence variation. The modified probe contains at least one artificial mismatch relative to the control nucleic acid target in addition to any mismatch(es) arising from the sequence variation. The invention has direct and advantageous application to numerous existing hybridization methods, including, applications that employ, for example, the Polymerase Chain Reaction, allele-specific nucleic acid sequencing methods, and diagnostic hybridization methods.

Abstract: The cystic fibrosis gene and its gene product are described for both the normal and mutant forms. The genetic and protein information is used in developing DNA diagnosis, protein diagnosis, carrier and patient screening, drug and gene therapy, cloning of the gene and manufacture of the protein, and development of cystic fibrosis affected animals.

Abstract: The present invention relates to an improved method for producing primed nucleic acid templates. Specifically, it relates to a method, compositions and kits therefor, of increasing the specificity of primer extension reactions by hybridizing primer to template in the presence of single-stranded nucleic acid binding protein.

Abstract: Methods are disclosed for producing chimeric nucleic acid molecules with two or more functionalities. A chimeric library is generated in which individual chimeric molecules combine the functions or characteristics of two or more parent libraries, each parent library having been selected through the SELEX procedure for a specific function or feature. The chimeric molecules of this invention are useful in a variety of ways, including having improved affinities for a target molecule, enhancing assembly of multi-component molecules, and promoting reactions between two molecules.

Abstract: A method for conducting sequential nucleic acid hybridization steps is described, whereby the ability of earlier-used primers or probes to participate in subsequent hybridization steps can be minimized, even though the differences between primer lengths are relatively small. It also relates to a rapid and quantitative method for the sequential synthesis of polynucleotide sequences by using a plurality of oligonucleotide primers, with the earlier utilized primers causing a minimum of interference with the subsequent primed synthesis reactions, yet without the need for intermediate purification steps.

Abstract: This invention pertains to methods for detecting mutations in the human ataxia telangiectasia (ATM) gene. The invention provides DNA sequences immediately flanking the exons in the 3' half of the gene. Also provided are primers that can be used in the polymerase chain reaction to amplify segments of the ATM gene corresponding to each of the 65 coding exons including its immediately flanking sequences. A number of mutations found in the human ATM gene are described also.

Abstract: The present invention relates to a method for the elimination of false negative test results in assays for the detection of amplified analyte nucleic acid in a sample. Prior to amplification, an internal control is added to the sample, comprising a nucleic acid distinguishable from the analyte nucleic acid, that can be amplified with the same amplification reagents as the analyte nucleic acid. Preferably the internal control comprises a nucleic acid sequence corresponding to the analyte nucleic acid that has been mutated to discriminate it from the analyte nucleic acid.

Abstract: Nucleic acid capture moieties, methods of using nucleic acid capture moieties, reaction mixtures including nucleic acid capture moieties, and kits including nucleic acid capture moieties are disclosed.

Abstract: An expression vector for the expression of an exogenous gene in an animal cell, which contains a chick .beta.-actin gene promoter and a restriction enzyme site for incorporating an exogenous gene at the downstream of said promoter and a process for expressing an exogenous gene, which comprises incorporating said exogenous gene into the expression vector at the restriction enzyme site for incorporating the exogenous gene, introducing said vector into an animal cell and culturing the obtained transformed animal cell. The expression system of the present invention can be applied to an expression of any exogenous gene and has an extremely high expression efficiency and is applicable to a wide range of host cells, and hence can sufficiently be utilized for industrial-scale production of a useful material.

Abstract: The invention provides nuclease protection assay comprising: (A) attaching a nucleic acid probe comprising a first nucleotide sequence to a solid surface area; (B) contacting the nucleic acid probe with a nucleic acid template under conditions that promote hybridization between complementary polynucleotides, forming a probe-template complex if the template includes a segment that is complementary to the probe; (C) contacting the probe-template complex with a nuclease effective to selectively cleave the nucleotide bonds of (1) the first nucleotide sequence when the first nucleotide sequence is single stranded or (2) mismatched regions of the first nucleotide sequence when the first nucleotide sequence is in duplex nucleic acid; and (D) detecting the presence of duplex nucleic acids formed by the probe and template nucleic acids by detecting the presence of the first nucleotide sequence.