Abstract: Methods for linearly amplifying mRNA to produce antisense RNA are provided. In the subject methods, mRNA is converted to double-stranded cDNA using a promoter-primer having a poly-dT primer site linked to a promoter sequence so that the resulting double-stranded cDNA is recognized by an RNA polymerase. The resultant double-stranded cDNA is then transcribed into antisense RNA in the presence of a reverse transcriptase that is rendered incapable of RNA-dependent DNA polymerase activity during this transcription step. The subject methods find use a variety of different applications in which the preparation of linearly amplified amounts of antisense RNA is desired. Also provided are kits for practicing the subject methods.

Abstract: A method for determining the activity of a nucleotide polymerizing enzyme in a sample, and use of the method for determining HIV 1 RT- and Herpes Simplex DNA-polymerase activity. The enzyme is captured by means of a nonoclonal antibody which is immobilized to a solid carrier and is capable of binding the enzyme without detrimentally effecting the enzyme activity. Contaminants and disturbing factors are removed and the nucleotide polymerization starts by the addition of a reaction solution containing a primer/template construct and nucleotides substrate, the reaction conditions being chosen such that they promote permanent association between antibody enzyme- and primer/template constructs. When necessary a nucleotide substrate, primer/template and reaction solution are washed away from the newly synthesized polymer, and the amount of nucleotide which as been incorporated into the polymer is determined, and the activity of the enzyme is determined with the guidance of this determination.

Abstract: The present invention provides a fast, simple and direct covalent bond formation between two strands of nucleotide sequences. Non-modified first strand nucleotide sequences are hybridized with second strand nucleotide sequences, of which certain specific base structure(s) is modified by chemical reagents in order to generate covalent bonding with the first strand. While the hybridization of these two strand nucleotide sequences generates double-stranded hybrid duplexes between their homologues, covalent bond formation occurs in the region of modified base-pairs. Since neither a polymerase chain restriction nor a restriction enzyme digestion can be performed with the covalently bonded hybrid duplexes, the present invention can be used to subtract common sequences during subtractive hybridization, to inhibit nonspecific contamination during subcloning and to increase binding stability of antisense probes during in situ hybridization as well as gene therapy.

Abstract: A set of contiguous and partially overlapping cDNA sequences and polypeptides encoded thereby, designated as PS108 and transcribed from prostate tissue, is described. These sequences are useful for the detecting, diagnosing, staging, monitoring, prognosticating, in vivo imaging, preventing or treating, or determining the predisposition of an individual to diseases and conditions of the prostate, such as prostate cancer. Also provided are antibodies which specifically bind to PS108-encoded polypeptide or protein, and agonists or inhibitors which prevent action of the tissue-specific PS108 polypeptide, which molecules are useful for the therapeutic treatment of prostate diseases, tumors or metastases.

Abstract: Compositions and methods are described for diagnosing periodontal disease, and in particular, early-onset periodontal disease. Nucleic acid-based testing is described which permits the detection of a high risk haplotype.

Abstract: Helicobacter pylori is known to cause or be a cofactor in type B gastritis, peptic ulcers, and gastric tumors. In both developed and developing countries, a high percentage of people are infected with this bacterium. The present invention relates generally to certain H. pylori proteins, to the genes which express these proteins, and to the use of these proteins for diagnostic and vaccine applications. Specifically, molecular cloning, nucteotide, and amino acid sequences for the H. pylori cytotoxin (CT), the "Cytotoxin Associated Immunodominant" (CAI) antigen, and the heat shock protein (hsp60). are described herein.

Abstract: The present invention is based at least in part on the discovery of the genomic structure of the human SR-BI gene and on the identification of polymorphic regions within the gene. Accordingly, the invention provides nucleic acids having a nucleotide sequence of an allelic variant of an SR-BI gene and nucleic acids having an SR-BI intronic sequence. The invention also provides methods for identifying specific alleles of polymorphic regions of an SR-BI gene, methods for determining whether a subject has or is at risk of developing a disease which is associated with a specific allele of a polymorphic region of an SR-BI gene, and kits for performing such methods.

Abstract: A method based on polymerase chain reaction (PCR) amplification and oligonucleotide ligase assay (OLA) reaction is provided for analyzing complex genetic systems in a single reaction vessel. The method involves simultaneously incubating a sample containing one or more target polynucleotides with PCR primers and OLA probes in a single reaction mixture. The presence of variant polynucleotide sequences in the sample is determined by detecting and identifying the products of the OLA reaction.

Abstract: Methods and compositions related to AIDS are disclosed. Using the methods of the present invention, candidate compounds may be screened for the ability to inhibit reverse transcriptase of human immunodeficiency virus ("HIV RT"). Active HIV RT mutants may be detected by the disclosed methods. The present invention also discloses methods for screening for compounds that inhibit HIV RT obtained from an individual patient. In another aspect, methods for testing the biological effectiveness of candidate compounds for the inhibition of HIV RT in vivo are disclosed.

Abstract: Detector nucleic acids are employed for detection of nucleic acid target sequences by fluorescence quenching mechanisms. The detector nucleic acid comprises at least two oligonucleotides and is partially single-stranded and partially double-stranded. One of the two dyes of a donor/acceptor dye pair is linked to the first oligonucleotide and the other is linked to a second oligonucleotide such that they are in close spatial proximity when the first and second oligonucleotides are base-paired and donor fluorescence is quenched. A single second oligonucleotide may be hybridized to the first oligonucleotide or multiple second oligonucleotides may be hybridized to the first oligonucleotide and to each other, forming a junction structure comprising multiple donor/acceptor dye pairs. The detector oligonucleotide retains its partially single-stranded and partially double-stranded conformation in the absence of target.

Abstract: The present invention provides a nucleic acid molecule encoding a CAULIFLOWER (CAL) gene product such as a nucleic acid molecule encoding Arabidopsis thaliana CAL and a nucleic acid molecule encoding Brassica oleracea CAL (BoCAL). The invention also provides a nucleic acid molecule encoding a truncated CAL gene product such as a nucleic acid molecule encoding Brassica oleracea var. botrytis CAL (BobCAL). The invention also provides a nucleic acid containing the Arabidopsis thaliana CAL gene, a nucleic acid molecule containing the Brassica oleracea CAL gene and a nucleic acid molecule containing the Brassica oleracea var. botrytis CAL gene. The invention further provides a kit for converting shoot meristem to floral meristem and a kit for promoting early flowering in an angiosperm. The invention provides a CAL polypeptide and an antibody that specifically binds CAL polypeptide.

Abstract: This invention discloses high-affinity oligonucleotide ligands to complex tissue targets, specifically nucleic acid ligands having the ability to bind to complex tissue targets, and the methods for obtaining such ligands. Tissue targets comprise cells, subcellular components, aggregates or cells, collections of cells, and higher ordered structures. Specifically, nucleic acid ligands to blood vessels are described.

Abstract: The invention relates to the detection of target nucleic acids or nucleic acid units in a sample, by obtaining a SER(R)S spectrum for a SER(R)S-active complex containing, or derived directly from, the target. The complex includes at least a SER(R)S-active label, and optionally a target binding species containing a nucleic acid or nucleic acid unit. In this detection method, the concentration of the target present in the SER(R)S-active complex, or of the nucleic acid or unit contained in the target binding species in the SER(R)S-active complex, is no higher than 10.sup.-10 moles per liter. Additionally or alternatively, one or more of the following features may be used with the method: i) the introduction of a polyamine; ii) modification of the target, and/or of the nucleic acid or nucleic acid unit contained in the target binding species, in a manner that promotes or facilitates its chemi-sorption onto a SER(R)S-active surface; iii) inclusion of a chemi-sorptive functional group in the SER(R)S-active label.

Abstract: Deletions in the EGF-R gene are found in many gliomas, breast tumors, and lung tumors. A particular truncated EGFR protein has been found in many tumors and provides diagnostic and therapeutic modalities.

Abstract: Methods of diagnosing primary congenital glaucoma, by detecting particular mutations in a human cytochrome P4501B1 (CYP1B1) gene, are disclosed. Methods include hybridization analysis, such as Southern or Northern analysis, which use hybridization of a mutant nucleic acid probe to the CYP1B1 gene; direct mutation analysis by restriction digest; sequencing of the CYP1B1 gene; hybridization of an allele-specific oligonucleotide with amplified genomic DNA; or identification of the presence of mutant proteins encoded by the CYP1B1 gene.

Abstract: Oligonucleotides in which one or more purine residues are substituted by pyrazolo[3,4-d]pyrimidines exhibit improved hybridization properties. Oligonucleotides containing pyrazolo[3,4-d]pyrimidine base analogues have higher melting temperatures than unsubstituted oligonucleotides of identical sequence. Thus, in assays involving hybridization of an oligonucleotide probe to a target polynucleotide sequence, higher signals are obtained. In addition, mismatch discrimination is enhanced when pyrazolo[3,4-d]pyrimidine-containing oligonucleotides are used as hybridization probes, making them useful as probes and primers for hybridization, amplification and sequencing procedures, particularly those in which single- or multiple-nucleotide mismatch discrimination is required.

Abstract: Hepatocyte culturing system, primer sets and an analytical method for selectively detecting and quantitatively assessing the levels of mRNA expression of the major isoenzymes of cytochrome P450 (CYP450 1A1, 1A2, 2B1/2, 2C11, 2E1, 3A1, 3A2 and 4A1), fatty acyl-CoA oxidase (FACO) and select Phase II conjugating enzymes (UDPGT, GST and ST) in the rat using specific 5' and 3' oligonucleotide primers and reverse transcriptase-polymerase chain reaction. The method closely reproduces the expression obtained from rat liver tissue following treatment with the same enzyme inducers. Constitutive and inducible expression was maintained by resuspending, culturing and then overlaying adult rat hepatocytes with an extracellular matrix such as Matrigel.RTM..

Abstract: A purified thermostable nucleic acid polymerase is obtained that has unique characteristics. Preferably the nucleic acid polymerase is DNA polymerase isolated from a Thermus aquaticus species and has a molecular weight of about 86,000-95,000 daltons. The thermostable nucleic acid polymerase may be native or recombinant and may be used in a temperature-cycling chain reaction wherein at least one nucleic acid sequence is amplified in quantity from an existing sequence with the aid of selected primers and nucleotide triphosphates. The nucleic acid polymerase is preferably stored in a buffer containing non-ionic detergents that lends stability to the nucleic acid polymerase. A preferred buffer contains glycerol, polyoxyethylated sorbitan monolaurate, ethoxylated nonyl phenol and gelatin.

Abstract: Nucleic acid probes substantially complementary to nucleic acid sequences within the bcl-1 locus are disclosed as well as polymerase chain reaction (PCR) primers substantially complementary to nucleic acid sequences within that locus that are useful in detecting t(11;14)(q13;q32) translocations associated with hematopoietic cancers. Further, the bcl-1 locus probes are useful in detecting bcl-1 amplifications found in about twenty percent of solid tumors, particularly in squamous cell and mammary carcinomas. Diagnostic/prognostic methods for cancer are disclosed, as well as cancer research methods using the bcl-1 probes of this invention.

Abstract: A method and apparatus is disclosed to reduce evaporation of solutions, particularly aqueous solutions, using a polymeric material such as polysucrose, polyvinylpyrrolidone and polyethylene glycol. A microscope slide and cover glass assembly utilizing the polymeric material in solution and method of use are described.

Abstract: The invention provides a diagnostic method and kits for SCA III syndrome. The method comprises attaching a portion of SCA III gene containing copies of 73 trinucleotide(CAG) repeat units to a substrate; amplifying a DNA segment containing copies of the trinucleotide repeat units from the genomic DNA of a testee using two labeled primers under the suitable condition for carrying out polymerase chain reaction (PCR); hybridizing the gene with the PCR product by amplifying DNA segment; and analyzing the results of the hybridization. A SCA III patient can be effectively diagnosed by examining the increase extent of the number of the TNR characteristic of the disease-associated gene with the aid of reverse dot hybridization technique or PCR-MPH.

Abstract: A nucleotide sequence encoding a katG/lacZ fusion protein is useful for assaying the enzymatic activity of the katG gene product. A process of selecting a compound that is toxic against an isoniazid-resistant mycobaterial strain comprises incubating a catalase peroxidase enzyme with an isoniazid to produce a compound that restores isoniazid susceptability to the isoniazid-resistant mycobaterial strain.

Abstract: Methods are described for the identification and preparation of high-affinity nucleic acid ligands to TGF.beta.. Included in the invention are specific RNA ligands to TGF.beta.1 identified by the SELEX method. Also included are RNA ligands that inhibit the interaction of TGF.beta.1 with its receptor.

Abstract: Disclosed are compositions and a method for amplification of nucleic acid sequences of interest. The method is based on stand displacement replication of the nucleic acid sequences of interest by multiple primers. In one preferred form of the method, referred to as multiple strand displacement amplification, two sets of primers are used, a right set and a left set. The primers in the right set are complementary to one strand of the nucleic acid molecule to be amplified and the primers in the left set are complementary to the opposite strand. The 5' end of primers in both sets are distal to the nucleic acid sequence of interest when the primers have hybridized to the nucleic acid sequence molecule to be amplified. Amplification proceeds by replication initiated at each primer and continuing through the nucleic acid sequence of interest. A key feature of this method is the displacement of intervening primers during replication by the polymerase.

Abstract: A method is disclosed for determining the presence of a polynucleotide analyte in a sample suspected of containing the analyte. The method comprises (a) forming as a result of the presence of an analyte a single stranded polynucleotide comprising a target polynucleotide binding sequence flanked by first and second polynucleotide sequences that differ from the sequence of the analyte or a sequence complementary to the analyte sequence, (b) forming multiple copies of the single stranded polynucleotide, and (c) detecting the single stranded polynucleotide. Also disclosed is a method of producing at least one copy of a single stranded polynucleotide.

Abstract: The invention relates to methods and compositions for simultaneously generating a plurality of polynucleotide sequencing ladders or PCR amplification products. Each sequencing ladder is generated from a recoverable primer, i.e., an oligonucleotide primer comprising a recovery tag. The recovery tag may be an oligonucleotide. Each sequencing ladder has a unique recovery tag. After the generation of the multiple sequencing ladders, the different sequencing ladders are separated from one another, i.e., purified, by binding to recovery tag binding compounds that have been immobilized on one or more solid supports. The recovery tag binding compounds are immobilized on the solid support in an addressable manner, i.e., the recovery tag binding compounds have distinct locations on the solid support. The binding of the sequencing ladders to the recovery tag binding compounds serves to separate the different polynucleotide sequencing ladders present in a given solution.

Abstract: A method of identification of differentially expressed messenger RNA (mRNA) which consists of synthesizing from a set of sequences of mRNA sets of fragments of complementary DNA (cDNA), which are separated with the aid of gel electrophoresis and the pictures of separation of the cDNA from different types of cells are compared and fragments with differential signal intensity are identified. For formation of the set of fragments the cDNA is cleaved with the aid of restriction nucleases. A method of cloning of differentially expressed mRNAs consists of synthesizing from sets of sequences of mRNAs from different types of cells sets of fragments of complementary DNA (cDNA) which are separated with the aid of gel electrophoresis, the pictures of the separation of the cDNA from different types of cells are compared, fragments of cDNA with different signal intensities are separated from the gel, amplified with the aid of a polymerase chain reaction and cloned to a plasmid or phage vector.

Abstract: An isothermal transcription based amplification assay for the detection or quantification of chemokine RNA uses primers and probes for sequences within the gene for RANTES, MIP-l.alpha. and MIP-1 .beta.. The quantitative system uses an internal control Q of a mutant version of each gene. Target specific primers and probes are also disclosed.

Abstract: A method for detecting mutations, such as a single base change or an addition or deletion of about one to four base pairs, is based on the use of an immobilized DNA mismatch-binding protein, such as MutS, which binds to a nucleic acid hybrid having a single base mismatch or unpaired base or bases, thereby allowing the detection of mutations involving as little as one base change in a nucleotide sequence. Such a method is useful for diagnosing a variety of important disease states or susceptibilities, including the presence of a mutated oncogene and the presence of DNA containing triplet repeat sequences which characterize several genetic diseases including fragile X syndrome. The present method is used to isolate or remove by affinity chromatography duplex DNA molecules containing mismatches such as error-containing molecules in PCR-amplified DNA samples. Methods for detecting and enriching minority sequences are disclosed.

Abstract: The present invention relates to a histidine kinase, two-component gene (CaHK1) from Candida albicans. CaHK1 encodes a 2471 amino acid protein with an estimated molecular mass of 281.8 kDa. Also provided are vectors, host cells, antibodies and recombinant methods for producing the same. The invention further relates agonists and antagonists and to screening methods for identifying agonists and antagonists of CaHK1 polypeptide activity. The invention additionally relates to diagnostic methods for detecting CaHK1 nucleic acids, polypeptides, and antibodies in a biological sample. The present invention further relates to novel antagonists and vaccines for the prevention or attenuation of infection by Candida albicans.

Abstract: Compositions and methods are provided for the modulation of ras expression. Oligonucleotides are provided which are targeted to nucleic acids encoding human ras. Oligonucleotides specifically hybridizable with mRNA encoding human H-ras, Ki-ras and N-ras are provided. Such oligonucleotides can be used for therapeutics and diagnostics as well as for research purposes. Methods are also disclosed for modulating ras gene expression in cells and tissues using the oligonucleotides provided, and for specific modulation of expression of activated ras. Methods for diagnosis, detection and treatment of conditions associated with ras are also disclosed.

Abstract: A method for detecting Byssochlamys nivea, Neosartorya fischeri and Zygossaccharomyces bailii micro-organisms by amplifying the genomic DNA of the targeted micro-organisms using as a primer a sequence contained on the internal transcript spacers (ITS) of the ribosomal unit, namely, for Byssochlamys nivea, on ITS1 corresponding to SEQ ID 1 and on ITS2 corresponding to SEQ ID 2; for Neosartorya fischeri, on ITS1 corresponding to SEQ ID 3, and on ITS2 corresponding to SEQ ID 4; and for Zygosaccharomyes bailii, on ITS1 corresponding to SEQ ID 5 and on ITS2 corresponding to SEQ ID 6.

Abstract: A method of identifying a peptide which permits or facilitates the transport of an active agent through a human or animal tissue. A predetermined amount of phage from a random phage library or preselected phage library is plated unto or brought into contact with a first side, preferably the apical side, of a tissue sample or polarized tissue cell culture. At a predetermined time, the phage which is transported to a second side of the tissue opposite the first side, preferably the basolateral side, is harvested to select transported phage. This modified phage is amplified in a host. This cycle of events is repeated (using the transported phage produced in the most recent cycle) a predetermined number of times to obtain a selected phage library containing phage which can be transported from the first side to the second side.

Abstract: The present invention provides labeled nucleic acid amplification oligonucleotides, which can be linear or hairpin primers or blocking oligonucleotides. The oligonucleotides of the invention are labeled with donor and/or acceptor moieties of molecular energy transfer pairs. The moieties can be fluorophores, such that fluorescent energy emitted by the donor is absorbed by the acceptor. The acceptor may be a fluorophore that fluoresces at a wavelength different from the donor moiety, or it may be a quencher. The oligonucleotides of the invention are configured so that a donor moiety and an acceptor moiety are incorporated into the amplification product. The invention also provides methods and kits for directly detecting amplification products employing the nucleic acid amplification primers. When labeled linear primers are used, treatment with exonuclease or by using specific temperature eliminates the need for separation of unincorporated primers.

Abstract: An improvement over the standard Sanger Method for nucleic acid sequencing is described. The novel method does not require denaturation of double-stranded template; rather, sequencing can be carried out directly on the double-stranded template. Embodiments are described with and without oligonucleotide primers.

Abstract: A method for detecting mutations, such as a single base change or an addition or deletion of about one to four base pairs, is based on the use of an immobilized DNA mismatch-binding protein, such as MutS, which binds to a nucleic acid hybrid having a single base mismatch or unpaired base or bases, thereby allowing the detection of mutations involving as little as one base change in a nucleotide sequence. Such a method is useful for diagnosing a variety of important disease states or susceptibilities, including the presence of a mutated oncogene and the presence of DNA containing triplet repeat sequences which characterize several genetic diseases including fragile X syndrome. The present method is used to isolate or remove by affinity chromatography duplex DNA molecules containing mismatches such as error-containing molecules in PCR-amplified DNA samples. Also provided are compositions and kits useful for practicing the methods of the present invention.

Abstract: Nucleic acids are made by adding a known nucleotide sequence to the 3' end of a first RNA having a known sequence at the 5' end to form a second RNA and reverse transcribing the second RNA to form a cDNA. In one embodiment, the first RNA is an amplified mRNA, the known sequence at the 5' end comprises a poly(T) sequence, the adding step comprises using a polyadenyltransferase to add a poly(A) sequence to the 3' end, the reverse transcribing step is initiated at a duplex region comprising the poly(T) sequence hybridized to the poly(A) sequence, the cDNA is converted to double-stranded cDNA by a polymerase initiating from a noncovalently joined duplex region, and the double-stranded cDNA is transcribed to form one or more third RNAs.

Abstract: A method for performing Gap-filling Ligase Chain Reaction (Gap-LCR) using an internal control which has been modified to contain a unique site for a restriction enzyme and which has approximately the same length and identical four LCR probe sites as the target nucleic acid.

Abstract: The present invention relates to materials and methods for identifying animals that are resistant or susceptible to diseases associated with intracellular parasites such as brucellosis, tuberculosis, paratuberculosis and salmonellosis. More particularly, the present invention relates to the identification of a gene, called NRAMP1, which is associated with the susceptibility or resistance of an animal, such as an artiodactyla to diseases such as brucellosis, tuberculosis, paratuberculosis and salmonellosis. Still more particularly, the present invention relates to the identification of specific sequences of bovine NRAMP1 which associate with resistance or susceptibility to ruminant brucellosis, tuberculosis, paratuberculosis and salmonellosis, and to the method of identifying said sequences to identify animals who are susceptible or resistant to disease.

Abstract: Methods and compositions are provided to obtain uniform amplification of nucleic acid templates having varied G+C contents by adding betaine and DMSO to the reaction mixture.

Abstract: The present invention provides new methods for preparing compositions for immunizing chickens against IBDV.

Abstract: A method of amplifying a mixture of different-sequence DNA fragments which may be formed from RNA transcription, or derived from genomic single- or double-stranded DNA fragments. The fragments are treated with terminal deoxynucleotide transferase and a selected deoxynucleotide, to form a homopolymer tail at the 3' end of the anti-sense strands, and the sense strands are provided with a common 3'-end sequence. The fragments are mixed with a homopolymer primer which is homologous to the homopolymer tail of the anti-sense strands, and a defined-sequence primer which is homologous to the sense-strand common 3'-end sequence, with repeated cycles of fragment denaturation, annealing, and polymerization, to amplify the fragments. In one embodiment, the defined-sequence and homopolymer primers are the same, i.e., only one primer is used. The primers may contain selected restriction-site sequences, to provide directional restriction sites at the ends of the amplified fragments.

Abstract: The invention relates to a method for diagnosing an individual as having atopy or a predisposition thereto. The method involves demonstrating in the individual the presence or absence of either: (i) an unusual variant form of the gene which codes for the beta sub-unit of the high affinity receptor for IgE (Fc.epsilon.RI.beta.), the variant form comprising a variation in exon (7) which encodes a glycine at amino acid residue 237 instead of glutamic acid which appears at residue 237 in individuals without atopy: or (ii) an unusual polymorphic form of the amino acid sequence for Fc.epsilon.RI.beta., the polymorphic form comprising glycine at amino acid residue 237 instead of glutamic acid which appears at residue 237 in individuals without atopy. Materials and other methods relating to the above diagnostic method are also described.

Abstract: A new class of nucleic acid compounds, referred to as nucleic acid ligands, have been shown to exist that have a specific binding affinity for three dimensional molecular targets, including cell surface macromolecules. The nucleic acid ligands are identified by the method of the invention referred to as the Systematic Evolution of Ligands by EXponential enrichment (SELEX), wherein a candidate mixture of nucleic acids are iteratively enriched and the high affinity nucleic acids are amplified for further partitioning. The high affinity nucleic acid ligands are useful in capturing target cells.

Abstract: The present invention describes a reaction chamber consisting of a reactant-containing aqueous solution, which may be in droplet form, coated with a hydrophobic powder such as polytetrafluoroethylene, polypropylene, polyolefins, polyethylene or hydrocarbon-coated particles such as hydrocarbon-coated silica. The polymer particles are preferably less than 500 microns in size. Charcoal, metal powder or silica powder can be inserted inside or on the surface of the reaction chamber droplet. Also, a reaction chamber droplet containing a first reactant such as an enzyme-bound bead can be combined with a second reaction chamber droplet containing a second reactant to mix the reactants in the two chambers resulting in a subsequent reaction. A dialysis or filtration membrane can also be placed on a reaction chamber droplet for dialysis using the droplet chamber. Electrochemical reactions can be performed by placing the droplet chamber between two electrodes and producing current flow between them.

Abstract: The present invention is directed to methods for resisting or promoting template independent nucleotide addition to the 3' terminus of a DNA duplex. The process comprises amplifying a target nucleic acid using primers which comprise a 5' terminal sequence which resists or promotes non-templated nucleotide addition to the 3' terminus of the complementary nucleic acid strand. The invention is also directed to a kit for cloning 3' nucleotidylated duplex DNA.

Abstract: The instant invention provides a means for quantitatively determining the vel of tumor suppressor gene p53 by determination of level of messenger ribonucleic acid (mRNA) of the gene in a sample when compared with a prepared standard. The assay is quantitative in that the specific number of copies of the p53 mRNA in a sample may be derived from a curve from a standard of p53 RNA. The RNA used in preparation of a standard curve to quantitate RNA is generated using a plasmid which is part of the invention. In the assay, the RNA is produced by a protein (RNA polymerase) that reads the DNA message and manufactures an RNA copy. The RNA content of the transcribed sample is determined spectrophotometrically to measure the molar concentration.

Abstract: A set of contiguous and partially overlapping RNA sequences and polypeptides encoded thereby, designated as PS112 and transcribed from prostate tissue is described. A fully sequenced clone representing a continuous sequence of PS112 is also disclosed. These sequences are useful for the detecting, diagnosing, staging, monitoring, prognosticating, preventing or treating, or determining the predisposition of an individual to diseases and conditions of the prostate, such as prostate cancer. Also provided are antibodies which specifically bind to PS112-encoded polypeptide or protein, and agonists or inhibitors which prevent action of the tissue-specific PS112 polypeptide, which molecules are useful for the therapeutic treatment of prostate diseases, tumors or metastases.

Abstract: Disclosed herein is a method for detecting a mismatch in a duplex nucleic acid, involving: a) contacting the duplex nucleic acid with a reactive agent under conditions which permit the agent to bind but not cleave a mismatch in said duplex nucleic acid; b) detecting binding of the agent to the duplex nucleic acid as an indication of the presence of a mismatch in the duplex nucleic acid; c) contacting the duplex nucleic acid with the reactive agent under conditions which permit the agent to cleave a mismatch in the duplex nucleic acid; and d) detecting a cleavage product as an indication of the presence of a mismatch in the duplex nucleic acid.

Abstract: A method of gene synthesis is disclosed. The gene synthesis method permits the codons of [m] natural gene to be changed to allow preferential transcription and translation of the synthetic gene in transgenic organisms. The method utilizes a combination of enzymatic and chemical synthesis of DNA and significantly reduces the cost, time and number of steps required for construction of synthetic genes.