Involving The Making Of Multiple Rna Copies Patents (Class 435/91.21)
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Patent number: 6498023Abstract: The present invention provides a method for the rapid simultaneous production of a plurality of single-stranded DNA circles having a predetermined size and nucleotide sequence using pre-designed hairpin oligonucleotides containing complementary sequences for directing ligation to form dumbbell-shaped monomers followed by heat denaturation to yield single-stranded DNA circles.Type: GrantFiled: November 28, 2000Date of Patent: December 24, 2002Assignee: Molecular Staging, Inc.Inventor: Patricio Abarzúa
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Patent number: 6492143Abstract: This invention provides novel libraries of olfactory receptor odorant/ligand-binding domains and methods of making and using them. The invention also provides libraries of vectors and cells comprising these nucleic acid constructs. The compositions and methods of the invention are used to identify novel ligand-binding domains for olfactory neuron odorant receptors and their ligands. Thus, the compositions and methods of the invention can be used to generate novel odorants, to screen for toxic odorants, or to manipulate an animal's olfactory response.Type: GrantFiled: December 17, 1999Date of Patent: December 10, 2002Assignee: The John Hopkins UniversityInventors: Randall R. Reed, King-Wai Yau, Dietmar Krautwurst
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Patent number: 6485911Abstract: The present invention describes the identification, isolation and cloning of two human presenilin genes, PS-1 and PS-2, mutations in which lead to Familial Alzheimer's Disease. Also identified are presenilin homologue genes in mice, C. elegans and D. melanogaster. Transcripts and products of these genes are useful in detecting and diagnosing Alzheimer's disease, developing therapeutics for treatment of Alzheimer's disease, as well as the isolation and manufacture of the protein and the constructions of transgenic animals expressing the mutant genes.Type: GrantFiled: August 11, 2000Date of Patent: November 26, 2002Assignees: HSC Research and Development Limited Partnership, The Governing Council of the University of TorontoInventors: Peter H. St. George-Hyslop, Johanna M. Rommens, Paul E. Fraser
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Patent number: 6485903Abstract: An oligonucleotide probe is disclosed, the probe including an oligonucleotide, a fluorescer molecule attached to a first end of the oligonucleotide and a quencher molecule attached to the opposite end of the oligonucleotide. The probe is rendered impervious to digestion by the 5′→3′ exonuclease activity of a polymerase and the 3′→5′ extension of by a polymerase. The invention also includes methods for performing combined PCR amplification and hybridization probing, one such method including the steps of contacting a target nucleic acid sequence with PCR reagents and an oligonucleotide probe as described above, and subjecting these reagents to thermal cycling. One preferred refinement of the above method further includes the addition of a strand displacer to facilitate amplification.Type: GrantFiled: April 3, 1997Date of Patent: November 26, 2002Assignee: PE Corporation (NY)Inventor: Paul E. Mayrand
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Patent number: 6472187Abstract: An object of the present invention is to establish a method for amplifying an RNA in a biologically-derived sample by preparing a reaction solution in which a nucleic acid synthesis is not inhibited even in the presence of various biologically-derived impurities and as well as to enable a convenient, rapid and highly sensitive analysis of the RNA in the sample. In a method of the present invention, a reaction solution having a pH higher than that of an ordinarily employed reaction solution and/or containing a polyamine and/or a sulfated polysaccharide.Type: GrantFiled: July 6, 2000Date of Patent: October 29, 2002Assignee: Shimadzu CorporationInventors: Hiroshi Tonoike, Naoyuki Nishimura
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Patent number: 6462255Abstract: A novel method of over expressing genes in plants is provided. This method is based on the RNA amplification properties of plus strand RNA viruses of plants. A chimeric multicistronic gene is constructed containing a plant promoter, viral replication origins, a viral movement protein gene, and one or more foreign genes under control of viral subgenomic promoters. Plants containing one or more of these recombinant RNA transcripts are inoculated with helper virus. In the presence of helper virus recombinant transcripts are replicated producing high levels of foreign gene RNA. Sequences are provided for the high level expression of the enzyme chloramphenicol acetyltransferase in tobacco plants by replicon RNA amplification with helper viruses and movement protein genes derived from the tobamovirus group.Type: GrantFiled: October 8, 1999Date of Patent: October 8, 2002Assignee: Large Scale Biology CorporationInventor: Thomas H. Turpen
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Publication number: 20020102603Abstract: The present invention relates to methods and compositions for the treatment and diagnosis of cardiovascular disease, including, but not limited to, atherosclerosis, ischemia/reperfusion, hypertension, restenosis, and arterial inflammation. Specifically, the present invention identifies and describes genes which are differentially expressed in cardiovascular disease states, relative to their expression in normal, or non-cardiovascular disease states, and/or in response to manipulations relevant to cardiovascular disease. Further, the present invention identifies and describes genes via the ability of their gene products to interact with gene products involved in cardiovascular disease. Still further, the present invention provides methods for the identification and therapeutic use of compounds as treatments of cardiovascular disease.Type: ApplicationFiled: August 11, 1999Publication date: August 1, 2002Inventors: DEAN A. FALB, MICHAEL A. GIMBRONE
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Patent number: 6423492Abstract: A method is disclosed for detecting single-stranded target nucleic acid (2) which comprises the steps of forming a hybrid between said target nucleic acid and a nucleic acid probe (4), said nucleic acid probe labelled with an enzyme reagent (6) which hydrolyses single-stranded nucleic acid but is substantially without effect on double-stranded nucleic acid, said hybrid formed under conditions of pH which are outside the activity range of said enzyme reagent, adjusting said pH to a value within the activity range of said enzyme reagent allowing said enzyme reagent substantially to hydrolyse any single-stranded nucleic acid present, and detecting said hybrid.Type: GrantFiled: October 14, 1999Date of Patent: July 23, 2002Assignee: Zetatronics LimitedInventor: Stuart Harbron
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Patent number: 6410227Abstract: Methods for screening pigs to determine which are more likely to produce larger litters and/or are less likely to produce larger litters are provided, based on identification of OPN alleles present in a sample of pig genomic DNA. Kits for use in such methods are also provided.Type: GrantFiled: December 12, 1997Date of Patent: June 25, 2002Assignee: Dalgety PLCInventors: John Alan Mileham, Graham Stuart Plastow, Olwen Irene Southwood
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Patent number: 6403313Abstract: The invention provides a homogeneous assay for nucleic acid hybridization. The fluorescent intensity of a hybridization medium containing a probe, a target and an intercalating agent is a function of the hybridization efficiency of the probe with respect to the target. The assay can detect specific hybridization between single-stranded probes and non-denatured double-stranded targets to form triplexes, thus obviating the need to denature the targets. The assay can also detect duplex hybridization complexes. The assay can be used to identify accessible regions in folded nucleotide sequences, to determine the number of mismatched pairs in a hybridization complex, and to map genomes.Type: GrantFiled: December 21, 1999Date of Patent: June 11, 2002Assignee: Ingeneus CorporationInventors: Jasmine I. Daksis, Pierre Picard, Glen H. Erikson
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Publication number: 20020064824Abstract: This invention relates to a method for producing a zinc finger nucleic acid binding protein comprising preparing a zinc finger protein according design rules, varying the protein at one or more positions, and selecting variants which bind to a target nucleic acid sequence by polysome display.Type: ApplicationFiled: May 8, 2001Publication date: May 30, 2002Inventors: Yen Choo, Michael Moore
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Patent number: 6395518Abstract: An oligonucleotide probe is disclosed, the probe including an oligonucleotide, a fluorescer molecule attached to a first end of the oligonucleotide and a quencher molecule attached to the opposite end of the oligonucleotide. The probe is rendered impervious to digestion by the 5′→3′ exonuclease activity of a polymerase and the 5′→3′ extension of by a polymerase. The invention also includes methods for performing combined PCR amplification and hybridization probing, one such method including the steps of contacting a target nucleic acid sequence with PCR reagents and an oligonucleotide probe as described above, and subjecting these reagents to thermal cycling. One preferred refinement of the above method further includes the addition of a strand displacer to facilitate amplification.Type: GrantFiled: May 19, 1997Date of Patent: May 28, 2002Assignee: The Perkin-Elmer CorporationInventor: Paul E. Mayrand
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Patent number: 6387620Abstract: Methods are provided for the production of nucleic acid ligands against target molecules using a procedure known as Transcription-free Systematic Evolution of Ligands by EXponential enrichment (Transcription-free SELEX). The Transcription-free SELEX method assembles nucleic acid ligands from fragments of synthetic nucleic acids by annealing those fragments to a complementary template, and then ligating the fragments together.Type: GrantFiled: July 28, 1999Date of Patent: May 14, 2002Assignee: Gilead Sciences, Inc.Inventors: Jonathan Drew Smith, Larry Gold
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Patent number: 6387617Abstract: A nucleic acid molecule is provided which is initially catalytically inactive but which can complex with a specific co-factor, e.g., a nucleic acid molecule or a non-nucleic acid molecule, to form a catalytically active nucleic acid molecule. The catalytically active nucleic acid molecule can be used to detect the presence of a non-nucleic acid co-factor or of a nucleic acid co-factor using the nucleic acid sequence of the present invention.Type: GrantFiled: February 25, 1999Date of Patent: May 14, 2002Assignee: Intelligene Ltd.Inventors: Nathan Asher, Yaron Tikochinsky, Andy Ellington
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Publication number: 20020025939Abstract: The invention relates to immunotherapy of a mammalian subject by exposing the immune response cells of the subject to a nucleic acid construct encoding at least one hCG immunogenic epitope or precursor thereof such that the nucleic acid construct is taken up and processed by the immune response cells. The invention further relates to compositions comprising such hCG-encoding nucleic acid constructs.Type: ApplicationFiled: December 17, 1999Publication date: February 28, 2002Inventor: PATRICK L. IVERSEN
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Patent number: 6309819Abstract: The present invention relates to a diagnostic system for the detection of Crohn's disease and ulcerative colitis by means of nucleotide amplification reactions, in situ or in vitro which detection means are specific for the measles virus and are capable of distinguishing between the “wild” type and the attenuated measles virus strains. The invention further provides a use in the manufacture of a medicament for the treatment of Crohn's disease and/or ulcerative colitis which medicament comprises a suitable vector carrying antisense RNA to specific viral proteins which are consequently inhibited from being expressed in the host.Type: GrantFiled: October 23, 1997Date of Patent: October 30, 2001Inventor: Andrew Jeremy Wakefield
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Patent number: 6294337Abstract: A method for sequencing a target DNA fragment in which along with amplification of the target DNA fragment, nucleic acid transcripts are generated using an RNA polymerase and the amplified target DNA fragments are used as templates in the presence of terminators for nucleic acid transcription reaction and the generated nucleic acid transcripts are analyzed, characterized in that the amplification of target DNA fragments and the generation of nucleic acid transcripts are carried out at a constant temperature is disclosed. The amplification of target DNA fragments and the generation of nucleic acid transcripts can be carried out around the room temperature. A DNA sequencing method using a novel method in which without using a thermo-resistant RNA polymerase, the amplification of target DNA fragments and generation of nucleic acid transcript can be carried out simultaneously in parallel is provided.Type: GrantFiled: August 29, 2000Date of Patent: September 25, 2001Assignee: RikenInventor: Yoshihide Hayashizaki
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Patent number: 6287768Abstract: Arrays of polymeric targets stably associated with the surface of a rigid solid support are provided. In the subject arrays, the polymeric targets are arranged at least according to size. The polymeric targets of the subject arrays are generally biopolymeric compounds, e.g. nucleic acids and proteins, where ribonucleic acids and proteins are the preferred polymeric targets. The subject arrays find use in a variety of different applications, and are particularly suited for use in high through gene expression analysis applications.Type: GrantFiled: March 3, 2000Date of Patent: September 11, 2001Assignee: Clontech Laboratories, Inc.Inventors: Alex Chenchik, Paul Siebert
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Patent number: 6258543Abstract: This invention is directed to methods for the quantitative measurement of specific gene expression levels in biological samples. In one embodiment, methods for the quantitative monitoring of gene expression without either co-amplification of an added template or use of an endogenous constitutive transcript are provided. The former involves a duplex amplification reaction in which a single set of primers is used to amplify both genomic DNA and expressed mRNA from the same gene sequence. These primers are targeted for sequences flanking the splice junction and intron sequences for the mRNA and DNA respectively. By their use, any suitable nucleic acid amplification technology yields mRNA and DNA amplimers which are distinguishable by length and sequence heterogeneity.Type: GrantFiled: September 17, 1999Date of Patent: July 10, 2001Assignee: Xtrana, Inc.Inventors: John C. Gerdes, Jeffrey M. Marmaro
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Patent number: 6251594Abstract: There is disclosed a cancer diagnostic method based upon DNA methylation differences at specific CpG sites. Specifically, the inventive method provides for a bisulfite treatment of DNA, followed by methylation-sensitive single nucleotide primer extension (Ms-SNuPE), for determination of strand-specific methylation status at cytosine residues.Type: GrantFiled: June 9, 1998Date of Patent: June 26, 2001Assignee: USC/Norris Comprehensive Cancer Ctr.Inventors: Mark L. Gonzalgo, Peter A. Jones, Gangning Liang
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Patent number: 6251590Abstract: The present invention is directed to a method for identifying and/or cloning within a biological sample alternatively spliced nucleic acid regions ocurring between two physiological conditions, comprising hybridizing RNA derived from a test condition with cDNA derived from the standard condition and further identifying and/or cloning nucleic acids corresponding to alternative forms of splicing.Type: GrantFiled: March 24, 1998Date of Patent: June 26, 2001Assignee: ExonHit Therapeutics S.A.Inventors: Fabien Schweighoffer, Laurent Bracco, Bruno Tocque
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Patent number: 6248520Abstract: Isolated nucleic acid molecules encoding Thyroid Receptor-Associated Proteins (TRAPS) are provided. TRAPS are members of protein complexes that bind to nuclear hormone receptors in a ligand-dependent manner so that the receptor, upon activation by a corresponding hormone, regulates the transcription of a particular gene. Also provided are methods of replicating and expressing such isolated nucleic acid molecules, pharmaceutical compositions comprising TRAPS, and methods of modulating gene expression via administration of therapeutically effective amounts of such pharmaceutical compositions.Type: GrantFiled: July 6, 1998Date of Patent: June 19, 2001Assignee: The Rockefeller UniversityInventors: Robert G. Roeder, Joseph D. Fondell, Chao Xingyuan, Mitsuhiro Ito
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Patent number: 6248534Abstract: The present invention provides a method of detecting RNA, comprising: (a) preparing cDNA to be measured and standard cDNA by reverse transcription of RNA as the subject of detection and standard RNA respectively; (b) adding different adapters respectively to the standard cDNA prepared at stepwise concentrations; (c) adding an adapter other than said adapters to the cDNA to be measured; (d) mixing and amplifying the adapter-tagged cDNAs obtained in the steps (b) and (c); (e) measuring the amount ratio of the cDNA measured to the standard cDNA; and (f) detecting said RNA from the measurement results.Type: GrantFiled: November 24, 1999Date of Patent: June 19, 2001Assignees: Unitech Co., Ltd.Inventors: Kikuya Kato, Ryou Matoba
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Patent number: 6232073Abstract: The present invention provides using a truncated WT1 gene transcript as a marker for detecting cancer in a subject. The method provides detecting the truncated WT1 gene transcript in a sample from the subject where the truncated gene transcript is characterized by an absence of a 101 base pair segment of intron 5 between nucleic acid positions −101 and −1. Positive detection of the truncated WT1 gene transcript indicates the presence of cancer. The invention provides a truncated WT1 gene transcript characterized by an absence of a 101 base pair segment of intron 5 between nucleic acid positions −101 and −1 and having a length of about two thousand base pairs. The truncated gene transcript is further characterized by containing at their five prime end sequences normally confined to the fifth intron of the WT1 gene, exons six through ten at their three prime end, and an overall length of approximately 2 kb.Type: GrantFiled: November 5, 1999Date of Patent: May 15, 2001Assignee: Virginia Commonwealth UniversityInventors: Joy L. Ware, Chavaboon Dechsukhum, Carleton T. Garrett
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Patent number: 6221584Abstract: The invention concerns a method for the detection of telomerase activity wherein (a) a sample to be tested is provided, (b) a first primer suitable as a telomerase substrate and nucleoside triphosphates are added and the reaction mixture is incubated under conditions under which a primer extension by the telomerase can take place, (c) an amplification of the extension product produced by the telomerase is carried out, (d) the amplification product produced in step (c) is immobilized on a solid phase and (e) the immobilized amplification product is detected qualitatively or/and quantitatively. Furthermore the invention concerns a suitable reagent kit for carrying out the method.Type: GrantFiled: December 29, 1997Date of Patent: April 24, 2001Assignee: Roche Diagnostics GmbHInventors: Thomas Emrich, Hermann Leying, Matthias Hinzpeter, Gerlinde Karl
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Patent number: 6214588Abstract: Eukaryotic RNA polymerase II holoenzymes that contain RNA polymerase II and one or more regulatory proteins are described. These holoenzymes selectively initiate transcription in vitro when supplemented with general transcription factors. The regulatory proteins act positively and negatively to regulate transcription initiation, at least in part, via functional interactions with RNA polymerase II.Type: GrantFiled: January 26, 1996Date of Patent: April 10, 2001Assignee: Whitehead Institute Biomedical ResearchInventors: Richard A. Young, Anthony J. Koleske, Craig M. Thompson, David M. Chao
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Patent number: 6210891Abstract: The present invention provides a method of identifying a base at a target position in a single-stranded sample DNA sequence wherein an extension primer, which hybridizes to the sample DNA immediately adjacent to the target position, is provided and the sample DNA and extension primer are subjected to a polymerization reaction in the presence of a deoxynucleotide or dideoxynucleotide, whereby the deoxynucleotide or dideoxynucleotide will only become incorporated and release pyrophosphate if it is complementary to the base in the target position. Release of pyrophosphate is detected enzymatically and pyrophosphate detection enzyme(s) are included in the polymerization step.Type: GrantFiled: July 6, 1999Date of Patent: April 3, 2001Assignee: Pyrosequencing ABInventors: Pål Nyren, Mathias Uhlen, Mostafa Ronaghi
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Patent number: 6210883Abstract: Compounds and methods for treating lung cancer are provided. The inventive compounds include polypeptides containing at least a portion of a lung tumor protein. Vaccines and pharmaceutical compositions for immunotherapy of lung cancer comprising such polypeptides, or DNA molecules encoding such polypeptides, are also provided, together with DNA molecules for preparing the inventive polypeptides.Type: GrantFiled: March 18, 1998Date of Patent: April 3, 2001Assignee: Corixa CorporationInventors: Steven G. Reed, Tong Tong Wang
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Patent number: 6210931Abstract: Circular RNAs may be synthesized by inserting DNA fragments into a plasmid containing sequences having the capability of spontaneous cleavage and self-circularization. Insertion of the DNA fragments allows RNAs of predetermined size to be constructed. In addition, a two-dimensional polyacrylamide gel electrophoresis system having a second dimension more highly cross-linked than the first dimension permits the separation and analysis as well as the precise sizing of both linear and circular RNAs produced by the synthetic method.Type: GrantFiled: November 30, 1998Date of Patent: April 3, 2001Assignee: The United States of America as represented by the Secretary of AgricultureInventors: Paul A. Feldstein, Robert A. Owens, Laurene Levy, John W. Randles
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Patent number: 6207373Abstract: Methods and apparatus are provided for the analysis and determination of the nature of repeat units in a genetic target. In one method of this invention, the nature of the repeat units in the genetic target is determined by the steps of providing a plurality of hybridization complex assays arrayed on a plurality of test sites, where the hybridization complex assay includes at least a nucleic acid target containing a simple repetitive DNA sequence, a capture probe having a first unique flanking sequence and n repeat units, where n=0,1,2 . . . , or fractions thereof, being complementary to the target sequence, and a reporter probe having a selected sequence complementary to the same target sequence strand wherein the selected sequence of the reporter includes a second unique flanking sequence and m repeat units, where m=0,1,2 . . . , or fractions thereof, but where the sum of repeat units in the capture probe plus reporter probe is greater than 0 (n+m>0).Type: GrantFiled: February 25, 1998Date of Patent: March 27, 2001Assignee: Nanogen, Inc.Inventors: Ronald G. Sosnowski, Eugene Tu
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Patent number: 6200753Abstract: A method for increasing the specificity of hybridization between a nucleic acid probe and a nucleic acid sequence to be detected, by addition of blocker molecules, which are complementary to the probe, raise of temperature in order to melt non-perfectly matched hybrids of probe and detected nucleic acid sequences, and lowering of the temperature again.Type: GrantFiled: February 27, 1998Date of Patent: March 13, 2001Assignee: Intelligene Ltd.Inventor: Asher Nathan
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Patent number: 6197554Abstract: The present invention provides a fast, simple and specific method for generating a complete full-length cDNA library from single cells. The first reverse transcription of intracellular mRNAs with an oligo(dT)n-promoter primer introduces a recognition site for following transcription of newly reverse-transcribed cDNAs. The poly-nucleotide tailing of above cDNAs in addition to aforementioned promoter region further forms binding templates for specific PCR amplification. After repeating the reverse transcription, transcription, reverse transcription and PCR procedure, we can multiply a single copy of mRNA to two billion folds by calculation based upon the comparison between the amount of a synthesized cDNA library and that of theoretically presumed mRNAs within a cell (0.1 pg). In conjunction with a cell fixation and permeabilization step, the complete full-length cDNA library can be directly generated from few single cells without mRNA degradation.Type: GrantFiled: November 20, 1998Date of Patent: March 6, 2001Inventors: Shi-Lung Lin, Cheng-Ming Chuong, Shao-Yao Ying
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Thermolabile uracil-DNA-glycosylas, process for its preparation and use for removing uracil from DNA
Patent number: 6187575Abstract: Thermolabile enzyme with uracil-DNA-glycosylase activity which is in particular characterized by a high degree of purity, short half-lives and a content of contaminating foreign activities of less than 2%, a process for its isolation as well as the use thereof to remove the base uracil from DNA and in particular from PCR products containing uracil. The enzyme is obtainable from gram-positive microorganisms such as e.g. Arthrobacter or Micrococcus.Type: GrantFiled: August 6, 1998Date of Patent: February 13, 2001Assignee: Roche Diagnostics GmbHInventors: Harald Sobek, Manfred Schmidt, Bruno Frey, Klaus Kaluza -
Patent number: 6174672Abstract: A method, and kit for carrying out the method, is provided for the non-radioactive and enzymatic detection of reverse transcriptase.Type: GrantFiled: February 10, 1998Date of Patent: January 16, 2001Assignee: Retro-Tech GmbH Gesellschaft fur Retrovirale TechnologieInventors: Ortwin Faff, Alois Gabriel Hisem