Patents Assigned to Ambion, Inc.
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Publication number: 20070238118Abstract: Methods for obtaining nucleic acid from nucleated cells are disclosed, wherein RNA and fragmented genomic DNA can be sequentially obtained from the same starting material are disclosed. In some embodiments, protein can also be obtained from the same starting material as the RNA and DNA are obtained. According to certain methods, whole blood or a blood fraction comprising nucleated cells is combined with a capture surface and at least some of the leukocytes are retained on the surface. The RNA is released from the retained leukocytes, then the capture surface is treated with a suitable nuclease or other DNA fragmenting agent to release DNA fragments. In certain embodiments, either the released RNA, the released DNA, or both the released RNA and the released DNA are employed in one or more molecular biology application.Type: ApplicationFiled: April 6, 2007Publication date: October 11, 2007Applicant: Ambion, Inc.Inventor: Marianna M. Goldrick
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Patent number: 7250270Abstract: The present invention concerns the methods and compositions for preparing a tissue section or biological sample, particularly to preserve RNA in the section or sample, by not exposing or contacting the sample or section to a solution that is composed of mostly water. Tissue sections can be fixed, stained, and dehydrated for subsequent manipulation, including laser capture microdissection (LCM) for further analysis using methods and/or compositions of the invention.Type: GrantFiled: June 13, 2003Date of Patent: July 31, 2007Assignee: Ambion, Inc.Inventors: Marianna Goldrick, Juanita C. Gonzales
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Publication number: 20070148174Abstract: Methods and compositions for inhibiting and/or inactivating nucleases by employing nuclease inhibitors are provided. The nuclease inhibitors comprise anti-nuclease antibodies and non-antibody nuclease inhibitors. The anti-nuclease antibodies of the present invention may be a polyclonal or monoclonal antibodies, and may be anti-ribonuclease antibodies, anti-deoxyribonuclease antibodies, or antibodies to non-specific nucleases. A preferred embodiment comprises at least two nuclease inhibitors, and is referred to as a nuclease inhibitor cocktail. In some specific embodiments, the invention concerns methods of performing in vitro translation comprising obtaining a first nuclease inhibitor, which inhibitor is further defined as an anti-nuclease antibody, and placing the anti-nuclease antibody in an in vitro translation reaction. In many cases, the in vitro translation reaction comprises at least one nuclease, which may be a ribonuclease, a deoxyribonuclease, or a nonspecific nuclease.Type: ApplicationFiled: December 6, 2006Publication date: June 28, 2007Applicant: AMBION, INC.Inventors: W. Kudlicki, Matthew Winkler, Brittan Pasloske
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Publication number: 20070032418Abstract: Lead compounds were obtained in a high throughput screen (HTS) of angiogenin (ANG; a potent inducer of angiogenesis) enzyme activity, an RNase. One lead was shown to delay appearance of tumors in an animal tumor system, and to reduce the number of animals having tumors. Several lead compound analogs were found to be even more potent inhibitors of ANG activity compared to the original leads, and two were also found to have greater affinity for ANG than for pancreatic RNase. Other embodiments disclose a method comprising obtaining a ribonuclease inhibitor and a composition; and admixing the ribonuclease inhibitor and the composition to form an admixture, wherein a ribonuclease that may be present in the admixture is inhibited.Type: ApplicationFiled: February 25, 2004Publication date: February 8, 2007Applicant: Ambion, IncInventors: Robert Shapiro, Jeremy Jenkins, Richard Kao, Gary Latham
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Patent number: 7163793Abstract: Methods and compositions for inhibiting and/or inactivating nucleases by employing nuclease inhibitors are provided. The nuclease inhibitors comprise anti-nuclease antibodies and non-antibody nuclease inhibitors. The anti-nuclease antibodies of the present invention may be a polyclonal or monoclonal antibodies, and may be anti-ribonuclease antibodies, anti-deoxyribonuclease antibodies, or antibodies to non-specific nucleases. A preferred embodiment comprises at least two nuclease inhibitors, and is referred to as a nuclease inhibitor cocktail. In some specific embodiments, the invention concerns methods of performing in vitro translation comprising obtaining a first nuclease inhibitor, which inhibitor is further defined as an anti-nuclease antibody, and placing the anti-nuclease antibody in an in vitro translation reaction. In many cases, the in vitro translation reaction comprises at least one nuclease, which may be a ribonuclease, a deoxyribonuclease, or a nonspecific nuclease.Type: GrantFiled: September 30, 2003Date of Patent: January 16, 2007Assignee: Ambion, Inc.Inventors: W. Antoni Kudlicki, Matthew M. Winkler, Brittan L. Pasloske
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Publication number: 20060228731Abstract: Compositions and method for making and using a synthetic bovine DNase I are disclosed. More particularly, the sbDNase I of the present invention is a versatile enzyme that cleaves DNA nonspecifically to release 5?-phosphorylated nucleotides. The sbDNase I molecules of the present invention find particular use in a wide range of molecular biology applications, including: degradation of contaminating DNA after RNA isolation; RNA clean-up prior to, or in conjunction with, RT-PCR after in vitro transcription; identification of protein binding sequences on DNA (DNase I footprinting); prevention of clumping when handling cultured cells; tissue dissociation and creation of fragmented DNA for in vitro recombination reactions.Type: ApplicationFiled: February 23, 2006Publication date: October 12, 2006Applicant: Ambion, Inc.Inventors: Gary Latham, Jon Kemppainen
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Patent number: 7067298Abstract: Compositions and method for making and using a synthetic bovine DNase I are disclosed. More particularly, the sbDNase I of the present invention is a versatile enzyme that cleaves DNA nonspecifically to release 5?-phosphorylated nucleotides. The sbDNase I molecules of the present invention find particular use in a wide range of molecular biology applications, including: degradation of contaminating DNA after RNA isolation; RNA clean-up prior to, or in conjunction with, RT-PCR after in vitro transcription; identification of protein binding sequences on DNA (DNase I footprinting); prevention of clumping when handling cultured cells; tissue dissociation and creation of fragmented DNA for in vitro recombination reactions.Type: GrantFiled: April 22, 2003Date of Patent: June 27, 2006Assignee: Ambion, Inc.Inventors: Gary Latham, Jon Kemppainen
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Patent number: 7033749Abstract: The present invention relates to nuclease resistant nucleic acids in general and ribonuclease resistant RNAs in particular. Methods of making and using such nucleic acids are disclosed.Type: GrantFiled: April 10, 2002Date of Patent: April 25, 2006Assignees: Ambion, Inc., Cenetron Diagnostics LLCInventors: Brittan L. Pasloske, Dwight B. DuBois, David M. Brown, Matthew M. Winkler
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Patent number: 6994963Abstract: This invention pertains to a method for generating a pool of nucleic acid fragments useful for in vitro recombination and the creation of novel DNA sequences that encode desirable proteins or enzymes. The invention provides a defined mixture of nucleic acids and methods for use in the synthesis, mutagenesis, and recombination of nucleic acids. Nucleic acids may be synthesized by creating a nucleic acid extension ladder, annealing the extension ladder to template nucleic acids, and further extending the ladder of nucleic acids. The invention also relates to methods for performing repeated cycles of synthesis for the purpose of mutagenesis or recombination, methods for producing mutant peptides and proteins from the mutagenized or recombined nucleic acids, and methods for selecting a peptide, polypeptide or protein having altered biological activities.Type: GrantFiled: July 10, 2000Date of Patent: February 7, 2006Assignee: Ambion, Inc.Inventors: George L. Murphy, Robert A. Setterquist, Andrew D. Ellington
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Publication number: 20050232934Abstract: The present invention relates to a gene that encodes a hyperactive reverse transcriptase having DNA polymerase activity and substantially reduced RNase H activity, vectors containing the gene and host cells transformed with the invention. The present invention also includes a method of producing the hyperactive reverse transcriptase, producing cDNA from mRNA using the reverse transcriptase of the invention, kits and assay templates made using the hyperactive reverse transcriptase.Type: ApplicationFiled: April 19, 2004Publication date: October 20, 2005Applicant: Ambion, Inc.Inventors: Liangjing Chen, Robert Setterquist, Gary Latham
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Patent number: 6897069Abstract: A system and method are described for electroporating a sample that utilizes one or more sets of electrodes that are spaced apart in order to hold a surface tension constrained sample between the electrodes. The first electrode is connected to the lower body of the system while the second electrode is connected to the upper body. Both electrodes are connected to a pulse generator. Each electrode has a sample contact surface such that the first electrode and the second electrode may be positioned to hold a surface tension constrained sample between the two sample contact surfaces and the sample may receive a selected electric pulse.Type: GrantFiled: June 8, 2004Date of Patent: May 24, 2005Assignee: Ambion, Inc.Inventors: Richard A. Jarvis, Mike W. Byrom, Dmitriy Ovcharenko
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Patent number: 6891032Abstract: The present invention is directed to compositions and methods for removal of nucleic acid probes from sample nucleic acids, particularly when the sample nucleic acids are attached to a solid support. The invention also concerns methods of stripping and reusing nucleic acid blots.Type: GrantFiled: February 25, 2002Date of Patent: May 10, 2005Assignee: Ambion, Inc.Inventors: David Brown, Matthew Winkler
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Patent number: 6825340Abstract: The present invention is a general method for inactivating or inhibiting ribonucleases. Ribonucleases are treated with a reducing agent and heat. RNA samples contaminated with ribonuclease may be treated with this method to protect them from degradation. The RNA may then be used directly in a variety of enzymatic reactions and molecular biology techniques. This method may also be applied to a variety of molecular biology reagents which may be contaminated with ribonuclease to protect an RNA from being degraded when incubated with the reagent.Type: GrantFiled: March 23, 2001Date of Patent: November 30, 2004Assignee: Ambion, Inc.Inventors: Brittan L. Pasloske, William Wu
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Publication number: 20040230048Abstract: The present invention provides methods and compositions, including kits, for the isolation and purification of mRNA, particularly poly(A) RNA. It concerns the use of isostabilizing salts such as TMAC and TEAC to reduce rRNA carryover during the purification process, thus facilitating the isolation of poly(A) RNA.Type: ApplicationFiled: May 11, 2001Publication date: November 18, 2004Applicant: AMBION, INC.Inventor: Richard C. Conrad
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Publication number: 20040219529Abstract: Compositions and method for making and using a synthetic bovine DNase I are disclosed. More particularly, the sbDNase I of the present invention is a versatile enzyme that cleaves DNA nonspecifically to release 5′-phosphorylated nucleotides. The sbDNase I molecules of the present invention find particular use in a wide range of molecular biology applications, including: degradation of contaminating DNA after RNA isolation; RNA clean-up prior to, or in conjunction with, RT-PCR after in vitro transcription; identification of protein binding sequences on DNA (DNase I footprinting); prevention of clumping when handling cultured cells; tissue dissociation and creation of fragmented DNA for in vitro recombination reactions.Type: ApplicationFiled: April 22, 2003Publication date: November 4, 2004Applicant: Ambion, Inc.Inventors: Gary Latham, Jon Kemppainen
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Patent number: 6812341Abstract: The present invention provides methods and compositions, including kits, for the isolation and purification of mRNA, particularly poly(A) RNA. It concerns the use of isostabilizing salts such as TMAC and TEAC to reduce rRNA carryover during the purification process, thus facilitating the isolation of poly(A) RNA.Type: GrantFiled: May 11, 2001Date of Patent: November 2, 2004Assignee: Ambion, Inc.Inventor: Richard C. Conrad
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Patent number: 6777210Abstract: The present invention is a general method for irreversibly inactivating ribonucleases. Ribonucleases are completely inactivated by treating them with a reducing agent and heat. RNA samples contaminated with ribonuclease may be treated with this method to protect them from degradation. The RNA may then be used directly in a variety of enzymatic reactions and molecular biology techniques. This method may also be applied to a variety of molecular biology reagents which may be contaminated with ribonuclease to protect an RNA from being degraded when incubated with the reagent.Type: GrantFiled: September 24, 1998Date of Patent: August 17, 2004Assignee: Ambion, Inc.Inventors: Brittan L. Pasloske, William Wu
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Publication number: 20040033602Abstract: The present invention concerns methods and compositions involving RNase III and polypeptides containing RNase III domains to generate RNA capable of triggering RNA-mediated interference (RNAi) in a cell. In some embodiments, the RNase III is from a prokaryote. RNase III activity will cleave a double-stranded RNA molecule into short RNA molecules that may trigger or mediate RNAi (siRNA). Compositions of the invention include kits that include an RNase III domain-containing polypeptide. The present invention further concerns methods using polypeptides with RNase III activity for generating RNA molecules that effect RNAi, including the generation of a number of RNA molecules to the same target.Type: ApplicationFiled: June 12, 2003Publication date: February 19, 2004Applicant: Ambion, Inc.Inventors: Lance P. Ford, David Brown
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Patent number: 6664379Abstract: Methods and compositions for inhibiting and/or inactivating nucleases by employing nuclease inhibitors are provided. The nuclease inhibitors comprise anti-nuclease antibodies and non-antibody nuclease inhibitors. The anti-nuclease antibodies of the present invention may be a polyclonal or monoclonal antibodies, and may be anti-ribonuclease antibodies, anti-deoxyribonuclease antibodies, or antibodies to non-specific nucleases. A preferred embodiment comprises at least two nuclease inhibitors, and is referred to as a nuclease inhibitor cocktail. In some specific embodiments, the invention concerns methods of performing in vitro translation comprising obtaining a first nuclease inhibitor, which inhibitor is further defined as an anti-nuclease antibody, and placing the anti-nuclease antibody in an in vitro translation reaction. In many cases, the in vitro translation reaction comprises at least one nuclease, which may be a ribonuclease, a deoxyribonuclease, or a nonspecific nuclease.Type: GrantFiled: September 25, 2000Date of Patent: December 16, 2003Assignee: Ambion, Inc.Inventors: W. Antoni Kudlicki, Matthew M. Winkler, Brittan L. Pastoske
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Patent number: 6586219Abstract: The present invention relates to compositions and methods for increasing the yields of polynucleotide synthetic reactions. In particular, it relates to an improved reaction mixture for use in in vitro RNA trancription and in various other enzymatic reactions in which a polynucleotide is synthesized. The reaction mixture uses high concentrations of total nucleotides, in the order of 12 mM to 40 mM, i.e. levels that were previously thought to be inhibitory. Other useful modifications are also disclosed.Type: GrantFiled: September 28, 2001Date of Patent: July 1, 2003Assignee: Ambion, Inc.Inventors: Susan Milburn, Marianna Goldrick, Matthew Winkler