Abstract: Patterns of pre-formed hybridizable nucleic acid oligomers are formed upon a substrate. The substrate is coated with molecules, such as aminosilanes, whose reactivity with nucleic acid molecules can be transformed by irradiation. The coated substrate exposed to patterned irradiation then contacted with pre-formed nucleic acid oligomers. The binding of the preformed nucleic acid oligomers to the coating molecules may be covalent or non-covalent (for example, ionic bonding or hydrogen bonding). If desired, a heterobifunctional crosslinker may be employed, before or after irradiation, with the coating to promote covalent binding of the nucleic acid oligomers to the coating molecules. Also, the irradiation step may be performed with the assistance of a positive-tone or negative-tone photoresist.

Abstract: Provided is an isolated double-stranded nucleic acid consisting essentially of the nucleotide sequences defined in the Sequence Listing by SEQ ID Nos:5-9. These are the ITS2 sequences for C. albicans, C. parapsilosis, C. tropicalis, C. glabrata and C. krusei. A method of diagnosing systemic candidiasis in a subject is also provided. The method comprises the steps of: (a) collecting blood from the subject into tubes containing detergent, polypropylene glycol, sodium poyantholesulfonate, and sodium ethylene diamine tetraacetic acid; (b) lysing Candida cells using ZYMOLYASE.RTM.-100T.TM.

Abstract: The present invention relates to materials and methods for diagnosing breast cancer in humans. It is based, at least in part, on the discovery that a substantial percentage of human breast cancer tissue samples contained nucleic acid sequences corresponding to a portion of the mouse mammary tumor env gene. In contrast, such sequences were absent in almost all other human tissues tested.

Abstract: The invention provides an apparatus for performing a process for amplification of specific nucleic acid sequences based upon the separation of nucleic acid strands by an electromagnetic field. This means of separation allows the use of mesophilic polymerases in the amplification process, thereby increasing the speed and fidelity of the amplification process, as well as the size of target nucleic acid that can be amplified.

Abstract: Disclosed is a gene situated in the region of the neuroblastoma consensus deletion 1p36.2-p36.1 which codes for a helix-loop-helix protein with the designation HEIR-1. The loss of this gene is significantly correlated with allelic tumor deletions in neuroblastomas and expression of this correlates inversely both N-myc overexpression in tumors and with N-myc expression in normal development. The cDNA and antibodies coding for HEIR-1 are used for the diagnosis of pathological conditions associated with aberration in the region of the neuroblastoma consensus deletion.

Abstract: A method is provided for detecting a target nucleic acid analyte in a sample. The method involves preparing an immobilized capture oligonucleotide that is complementary to the target analyte using a magnetic cycling method, incubating the sample with the immobilized capture oligonucleotide to capture the target analyte thereby forming a capture oligonucleotide-analyte complex, and detecting the presence of the capture oligonucleotide-analyte complex.

Abstract: Canavan disease, an autosomal recessive leukodystrophy, is caused by deficiency of aspartoacylase and accumulation of N-acetylaspartic acid in brain. Human aspartoacylase (ASP) cDNA spanning 1,435 bp has been cloned and expressed in E. coli. A base change, a854>c, has been found in 85% of the 34 Canavan alleles tested so far, which results in a missense glu285>ala mutation that is predicted to be part of the catalytic domain of aspartoacylase. Several additional mutations have also been identified. The invention therefore provides nucleic acid sequences, genes, polypeptides, antibodies, vectors containing the gene, host cells transformed with vectors containing the gene, animal models for the disease, methods for expressing the polypeptide, genetic screening methods and kits, diagnostic methods and kits, methods of treating Canavan disease and methods of genetic therapy for the disease.

Abstract: A nucleotide probe complex which enhances the ability to discriminate low level samples in electrochemiluminescent assays. The complex is composed of a platform molecule to which multiple copies of an organometallic electrochemiluminescent label and an oligonucleotide probe are separately attached. Preferably the complex is capped with streptavidin. Use of the complex permits detection of 1000 copies of analyte per sample in less than one hour.

Abstract: A diagnostic method for detecting a base pair mismatch in a DNA duplex, comprising the steps of contacting at least one strand of a first DNA molecule with the complementary strand of a second DNA molecule under conditions such that base pairing occurs contacting a DNA duplex potentially containing a base pair mismatch with a mispair recognition protein under conditions suitable for the protein to form a specific complex only with the DNA duplex having a base pair mismatch, and not with a DNA duplex lacking a base pair mismatch, and detecting any complex as a measure of the presence of a base pair mismatch in the DNA duplex.

Abstract: A method for sequencing a strand of DNA, including the steps of: providing the strand of DNA; annealing the strand with a primer able to hybridize to the strand to give an annealed mixture; incubating the mixture with a deoxyribonucleoside triphosphate, a DNA polymerase, and a chain terminating agent under conditions in which the polymerase causes the primer to be elongated to form a series of DNA products differing in length of the elongated primer, each DNA product having a chain terminating agent at its elongated end; the number of each DNA product being approximately the same for substantially all DNA products differing in length from 1 to 20 bases.

Abstract: An efficient method for sequencing large fragments of DNA is described. A subclone path through the fragment is first identified; the collection of subclones that define this path is then sequenced using transposon-mediated direct sequencing techniques to an extent sufficient to provide the complete sequence of the fragment.

Abstract: The present invention describes the formation of RecA protein catalyzed double-stranded probe:duplex linear target DNA complexes that are stable to deproteinization. The uses of this stable probe:target complex in diagnostic/DNA detection systems in in vitro and in situ DNA hybridization reactions is discussed. The probe:target complexes are also useful for diagnostic application in RecA protein facilitated DNA amplification reactions.

Abstract: Isolated polynucleotide molecules, and peptides encoded by these molecules, can be used in the analysis of alloantigen phenotypes, as well as in diagnostic and therapeutic applications relating to human platelet Pl.sup.A polymorphism. In this vein, a method for typing blood cell and platelet membrane glycoproteins entails an analysis of amplified cDNA, encoded by platelet and red blood cell mRNA.

Abstract: Compositions and methods for covalently immobilizing an oligonucleotide onto a polymer-coated solid support or similar structure are provided. Specifically, the polymer-coated support, such as a bead, possesses a large number of activatable moieties, preferably primary and secondary amines. An oligonucleotide is activated with a monofunctional or multifunctional reagent, preferably the homotrifunctional reagent cyanuric chloride. The resultant covalently immobilized oligonucleotides on the support serve as nucleic acid probes, and hybridization assays can be conducted wherein specific target nucleic acids are detected in complex biological samples. The beads or similar structures can be employed free in solution, such as in a microtiter well format; in a flow-through format, such as in a column; or in a dipstick. Additionally, dichlorotriazine oligonucleotides and processes for activating oligonucleotides by treatment with cyanuric chloride and derivatives are included in the present invention.

Abstract: DNA sequences and corresponding amino acid sequences from the HLA class II beta region of the human genome that are associated with insulin-dependent diabetes mellitus (IDDM) and Pemphigus vulgaris (PV) have been identified. Specifically, marker DNA sequences which detect either directly or indirectly the identity of the codon encoding for the amino acid at position 57 of the DQ.beta. protein sequence are disclosed as well as sequences from the DR.beta. region. These sequences may be used to generate DNA hybridization probes and antibodies for assays to detect a person's susceptibility to autoimmune diseases, such as IDDM and PV. Such antibodies and peptides encoded by said DNA sequences can be used therapeutically or prophylactically.

Abstract: Methods for covalent attachment of oligonucleotides to solid supports such that substantially all of the oligonucleotides are attached via their 5'-ends are provided. The solid supports with attached oligonucleotides are produced. Thiol-oligonucleotides are attached to bromoacetyl-derivatized polyacrylamide supports, or conversely, bromoacetyl-oligonucleotides are immobilized on thiol-polyacrylamide supports. In a further aspect, bromoacetyl-derivatized oligonucleotides, and polyacrylamide supports with linked oligonucleotides produced by coupling bromoacetyl-derivatized oligonucleotides with thiol-derivatized polyacrylamide solid supports or by coupling thiol-derivatized oligonucleotides with bromoacetyl-derivatized polyacrylamide supports as well as methods for capture of nucleic acids by oligonucleotides attached to polyacrylamide solid supports, either by direct capture or in sandwich hybridization formats are provided.

Abstract: A method for determining virus replication in human cells by human retrovirus using RNA amplification comprising detecting the hybridization of an RNA probe which specifically hybridizes with spliced RNA and not with genomic RNA. This method permits early detection of RNA replication resulting from primary infection without detecting non-replicating virus.

Abstract: A method for determining the presence and/or concentration of a target substance e.g. protein, nucleic acid, bioparticle etc. in a fluid sample is provided. The method disclosed combines elements of immunoassays, coated cup assays and magnetic particle separation to effect the quantitation and recovery of an analyte in solution. Also the method ensures the non-reorientation of magnetically collected material by linking the magnetic particles to a collection surface via a specific binding pair. This linkage immobilizes the magnetic-analyte-containing material and thus allows for vigorous washing and reagent addition without significant redistribution or displacement. Thus the assay of this invention offers the speed of diffusion controlled kinetics as in a ferrofluid assay, the speed of collection of labeled target substance as in a magnetic assay as well as the ability to magnetically monolayer the ferrofluid, all of which is combined with the ease of washing and signal detection found in a coated cup assay.

Abstract: The instant invention provides for the identification, diagnosis, monitoring, and treatment of invasive cells using the laminin 5 gamma-2 chain protein or nucleic acid sequence, or antibodies thereto.

Abstract: A process for producing a particular nucleic acid sequence from a given sequence of DNA or RNA in amounts which are large compared to the amount initially present, using PNAs in conjunction with the polymerase chain reaction (PCR) is disclosed. The process may be used to amplify relatively long DNA or RNA sequences of about 600 base pairs or longer, while reducing the generation of anomalous products and the tendency to preferentially amplify smaller nucleic acid (DNA or RNA) fragments when present in the heterozygous state with a larger allele.