Removing Nucleic Acid From Intact Or Disrupted Cell Patents (Class 435/270)
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Publication number: 20080261293Abstract: The present invention provides a method for the isolation of sperm DNA from swabs taken from rape victims without having to perform a change in buffers. Non-sperm cells from the victim are digested with an enzyme and solubilized, and then in the same buffer an enzyme capable of digesting soluble DNA is added and the victim's DNA is degraded, leaving only the rapist's DNA intact. Since no change of buffer is needed, no centrifugation or filtration steps are needed. The inventive method has utility particularly in the forensic science field.Type: ApplicationFiled: January 11, 2008Publication date: October 23, 2008Inventor: Alex Garvin
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Publication number: 20080261292Abstract: This invention relates to the use of tumor-derived or associated extracellular ribonucleic acid (RNA) found circulating in the plasma or serum fraction of blood for the detection, monitoring, or evaluation of cancer or premalignant conditions. Extracellular RNA may circulate as non-bound RNA, protein-bound RNA, lipid-RNA complexes, lipoprotein (proteolipid)-RNA complexes, protein-RNA complexes including within or in association with ribonucleoprotein complexes, nucleosomes, or within apoptotic bodies. Any intracellular RNA found in plasma or serum can additionally be detected by this invention. Specifically, this invention enables the extraction of circulating RNA from plasma or serum and utilizes nucleic acid amplification assays for the identification, detection, inference, monitoring, or evaluation of any neoplasm, benign, premalignant, or malignant, in humans or other animals, which might be associated with that RNA.Type: ApplicationFiled: April 24, 2007Publication date: October 23, 2008Applicant: ONCOMEDX, INC.Inventor: Michael S. Kopreski
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Publication number: 20080248559Abstract: A method for selectively separating and purifying RNA from a mixture solution of nucleic acid containing DNA and RNA, wherein the method comprising the steps of: (1-a) adsorbing nucleic acid; (1-b) washing; (1-c) subjecting to a DNase treatment; (1-d) washing; and (1-e) desorbing the RNA from a nucleic acid-adsorbing porous membrane by a recovering solution, wherein in the step (1-c), a total amount of a DNase solution is 130 ?l or less per 1 cm2 of the membrane. And a method for selectively separating and purifying RNA or DNA, which comprises the steps of: (2-a) adsorbing nucleic acid; (2-b) washing by a washing solution; and (2-c) desorbing the nucleic acid from a nucleic acid-adsorbing porous membrane, wherein the washing solution contains a water-soluble organic solvent having a concentration of 50% by weight or less, and does not contain a chaotropic salt.Type: ApplicationFiled: March 25, 2005Publication date: October 9, 2008Inventors: Hiroko Inomata, Rie Iwata
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Publication number: 20080241845Abstract: Methods are provided for isolation of chromatin fractions of nucleoproteins containing histone H1, H2A, H2B, H3 and H4 proteins and/or histone H1, H2A, H2B, H3 and/or H4 proteins, from intact cells. The methods preserve original patterns of covalent modifications of the histone proteins.Type: ApplicationFiled: April 2, 2008Publication date: October 2, 2008Inventors: Pedro Rodriguez-Collazo, Sanford Harrison Leuba, Jordanka Zlafanova
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Patent number: 7429470Abstract: An apparatus and method for purification of nucleic acids of cells or viruses are provided. The nucleic acid purification apparatus includes: a cell lysis capillary having a sample inlet through which samples and magnetic beads are introduced; a vibrator attached to the capillary and mixing the samples and the magnetic beads in the capillary; a laser generator attached to the capillary and supplying a laser to the capillary; and a magnetic force generator attached to the capillary and fixing the magnetic beads to a capillary wall. According to the method and apparatus, PCR yield can be increased since PCR inhibitors can be readily removed by means of a phase separation in a capillary. The use of an electromagnet ensures the removal of the PCR inhibitors. In addition, since cell lysis and DNA purification process can be simultaneously performed, LOC steps can be reduced.Type: GrantFiled: November 2, 2005Date of Patent: September 30, 2008Assignee: Samsung Electronics Co., Ltd.Inventors: Jeong-gun Lee, Young-nam Kwon, Young-a Kim
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Publication number: 20080233632Abstract: The present invention relates to a process for labeling and purification of nucleic acids of interest present in a biological sample to be treated, comprising: taking a single reaction vessel, introducing into the reaction vessel: the biological sample, at least one labeling reagent for nucleic acids, at least one solid support enabling the adsorption of said nucleic acids, any ingredient necessary for the labeling of the nucleic acids and/or for the immobilization of said nucleic acids on the support, incubating the contents of the reaction vessel, and isolating the nucleic acids thus labeled. The invention finds a preferred application in the diagnostics field.Type: ApplicationFiled: July 21, 2005Publication date: September 25, 2008Applicant: BIOMERIEUXInventors: Ali Laayoun, Lionel Menou, Frederic Ginot
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Publication number: 20080213870Abstract: The present invention provides a method for obtaining modified DNA from a biological specimen by obtaining a cell suspension from the specimen, if necessary; passing the cell suspension through a first filter under conditions sufficient to obtain filter-bound cells and suspended DNA; lysing the filter-bound cells under conditions sufficient to release cellular DNA; modifying the DNA bound to the filter under conditions sufficient to release the modified DNA from the filter into a flow-through volume; passing the flow-through volume through a second filter under conditions sufficient to capture the modified DNA to the second filter; and eluting the modified DNA from the second filter.Type: ApplicationFiled: March 3, 2008Publication date: September 4, 2008Inventors: Sean Wuxiong Cao, Jadwiga Katarzyna Markiewicz, George A. Green
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Publication number: 20080206746Abstract: Apparatus, reagents, and methods for isolating plasmid DNA from bacteria by alkaline lysis using a solid or immobilized P2 and/or P3 reagent in combination with a DNA-binding matrix.Type: ApplicationFiled: July 22, 2006Publication date: August 28, 2008Inventor: Xiyu Jia
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Publication number: 20080193912Abstract: Methods and compositions for releasing the nucleic acids from a variety of different types of microorganisms are provided. The method relies on a simplified lysis procedure that can be applied to many types of bacteria and fungal cells and is readily automated for high throughput screening methods. The method utilizes a high concentration of a chelating agent and a mixture of lysing enzymes to accomplish the disruption of microbial cell walls and allow the release of the nucleic acid.Type: ApplicationFiled: August 3, 2005Publication date: August 14, 2008Inventors: Yiu-Lian Fong, Azita Tabrizi
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Publication number: 20080160511Abstract: A capture probe suitable for use with a method for isolating miRNAs. A method for isolating an miRNA of interest from a sample comprising the miRNA of interest comprising providing the capture probe. A method for identifying an miRNA of interest.Type: ApplicationFiled: August 18, 2006Publication date: July 3, 2008Inventors: Elliott P. Dawson, Kristie E. Womble
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Publication number: 20080138886Abstract: Methods of isolating clinical-grade plasmid DNA from manufacturing processes, including large-scale fermentation regimes, are disclosed which encompass alternatives to two core unit operations common to plasmid DNA purification processes. The novel upstream and downstream purification processes disclosed herein provide for reduced production costs and increase process robustness. Either or both of the purification processes disclosed herein may be used in combination with additional purification steps known in the art that are associated with DNA plasmid purification technology.Type: ApplicationFiled: January 27, 2006Publication date: June 12, 2008Inventors: Jason C. Murphy, David B. Boyd, Adam Joel Kristopeit, Russel Jackson Lander, Michael Albert Winters
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Publication number: 20080131955Abstract: The present invention relates to methods of separating target DNA from mixed DNA in a sample. In some embodiments, the target DNA is present in target organisms. In other embodiments, the target organisms may be viruses, bacteria, fungi or combinations thereof. In some embodiments the sample contains cells having nuclei. In some embodiments the cells are mammalian cells. In some embodiments, the separated target organisms are treated to release their DNA which can be recovered.Type: ApplicationFiled: November 28, 2007Publication date: June 5, 2008Applicant: CANON U.S. LIFE SCIENCES, INC.Inventor: Michele R. Stone
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Publication number: 20080131954Abstract: The present invention relates to methods of separating target DNA from mixed DNA in a sample. In some embodiments, the target DNA may be viral DNA, prokaryotic DNA, fungal DNA or combinations thereof. In some embodiments the mixed DNA includes target DNA and non-target DNA.Type: ApplicationFiled: November 28, 2007Publication date: June 5, 2008Applicant: CANON U.S. LIFE SCIENCES, INC.Inventor: Michele R. Stone
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Publication number: 20080118972Abstract: Provided is a method of isolating nucleic acid from cells using a single surface, wherein a compound represented by Formula 1 is bound to the surface. Also provided are an apparatus for isolation of nucleic acids, and a bead for isolating nucleic acids.Type: ApplicationFiled: March 29, 2007Publication date: May 22, 2008Applicant: SAMSUNG ELECTRONICS CO., LTD.Inventors: Chang eun YOO, Sung-young JEONG, Young-rok KIM
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Publication number: 20080113348Abstract: The present invention relates to a method for cleaning and isolating nucleic acids using cationic detergents with the general formula (I): Y+R1R2R3R4X? ??(I) where Y can represent nitrogen or phosphorus R1, R2, R3 and R4 can represent independently from one another an unbranched or branched C1-C20-alkyl residue, C3-C6-alkenyl residue, C3-C6-alkinyl residue and/or a C6-C20-aryl residue as well as a C6-C26-aralkyl residue, and X— can represent an anion of an inorganic or organic single or multi-basic acid.Type: ApplicationFiled: July 15, 2005Publication date: May 15, 2008Inventor: Thorsten Singer
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Publication number: 20080113356Abstract: A method for separating and purifying a nucleic acid comprising steps of: (1) adding a lysis solution to a biomaterial to prepare a sample solution containing a nucleic acid, and adding a water-soluble organic solvent or a solution containing a water-soluble organic solvent to the sample solution thereby preparing a sample solution containing the water-soluble organic solvent; (2) contacting the sample solution containing the water-soluble organic solvent with a solid phase thereby adsorbing the nucleic acid on the solid phase; (3) contacting a washing solution with the solid phase thereby washing the solid phase in a state where the nucleic acid is adsorbed on the solid phase; and (4) contacting a recovering solution with the solid phase thereby desorbing the nucleic acid from the solid phase, wherein, in the step (1), the water-soluble organic solvent or the solution containing the water-soluble organic solvent is added separately in at least two batches.Type: ApplicationFiled: February 2, 2006Publication date: May 15, 2008Applicant: FUJIFILM CORPORATIONInventors: Tasuku Sasaki, Hiroko Inomata, Rie Iwata
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Publication number: 20080090227Abstract: The present invention provides for a method of drying and spreading chromosomes from various biological samples to yield optimal chromosomal spreading. The method requires preparing a biological sample for treatment, providing a cytogenetic chamber capable of setting predetermined conditions, pre-testing a portion of the biological sample in the cytogenetic chamber, and finally treating the remaining biological sample. The method is useful to yield metaphase chromosomes that are small and rounded, with very few overlapping or scattered chromosomes. Furthermore, the method is uses restricted ranges of temperature and relative humidity to achieve consistent chromosomal spreading. The morphologies of the chromosomes are preserved in order to execute banding techniques at 550 bands and chromosomal analysis on high-resolution chromosomes.Type: ApplicationFiled: October 11, 2006Publication date: April 17, 2008Inventor: Philip D. Buchanan
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Patent number: 7351378Abstract: The present invention provides an apparatus for extracting nucleic acid from a specimen, including traditionally difficult specimens, in preparation for polymerase chain reaction (PCR) analysis. The apparatus includes: a shaker, a centrifugal separator, a particle supply mechanism for supplying particles of different sizes, a liquid chemical supply mechanism, a conveyance arm for transporting the specimen or tube-containing specimen, and a control means. The control means is designed to shake tubes containing large and small diameter particles and transport the tubes through a centrifuge in order to separate the nucleic acid-containing material from the remainder of the specimen and crush the material to extract the nucleic acid.Type: GrantFiled: September 27, 2002Date of Patent: April 1, 2008Assignee: Kabushiki Kaisha Yakult HonshaInventors: Tsuyoshi Osanai, Eiji Yamamoto, Katsuhiro Tomaru, Junji Fujimoto, Koichi Watanabe, Takahiro Matsuki, Yukiko Miyamoto, Ryuichiro Tanaka
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Publication number: 20080050774Abstract: The present invention relates to an isolated polynucleotide of the complete chromosome of Bacillus licheniformis. The present invention also relates to isolated genes of the chromosome of Bacillus licheniformis which encode biologically active substances and to nucleic acid constructs, vectors, and host cells comprising the genes as well as methods for producing biologically active substances encoded by the genes and to methods of using the isolated genes of the complete chromosome of Bacillus licheniformis.Type: ApplicationFiled: November 5, 2004Publication date: February 28, 2008Applicants: Novozymes A/S, Novozymes, Inc.Inventors: Randy Berka, Michael Rey, Preethi Ramaiya, Jens Tonne Andersen, Michael Dolberg Rasmussen, Peter Bjarke Olsen
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Patent number: 7318998Abstract: The present invention relates, in general, to tissue decellularization and, in particular to a method of treating tissues, for example, heart valves, tendons and ligaments, so as to render them acellular and thereby limit mineralization and/or immunoreactivity upon implementation in vivo.Type: GrantFiled: March 24, 2003Date of Patent: January 15, 2008Assignee: Cryolife, Inc.Inventors: Steven Goldstein, Kirby S. Black
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Patent number: 7282475Abstract: The present invention provides lysis reagents, containers, methods and kits relating to the extraction or the extraction and isolation of a cellular component from a host cell. More specifically, the invention provides combinations of zwitterionic compounds that may be employed to aide in the extraction or the extraction and isolation of a cellular component from a host cell.Type: GrantFiled: August 23, 2005Date of Patent: October 16, 2007Assignee: Sigma-Aldrich Co.Inventors: Jeff Porter, Richard Mehigh
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Patent number: 7282361Abstract: The present invention provides a method and apparatus for to conduct transgenic and targeted mutagenesis screening of genomic DNA. This invention also provides a system for screening DNA for a designated genetic sequence. The system includes a computer having a processor, memory and web browser, wherein the computer receives instructions concerning the designated genetic sequence and other screening parameter selected from a remote user via a form of electronic communication, and an automatic screening device that analyzes samples of genomic DNA for the designated sequence.Type: GrantFiled: March 8, 2005Date of Patent: October 16, 2007Assignee: Transnetyx, Inc.Inventor: Timothy A. Hodge
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Publication number: 20070238114Abstract: Provided herein is a method and apparatus for disrupting cells and purifying nucleic acids in a single chip. The method comprises irradiating a chip with a laser beam, wherein the chip comprises a solid support on which a cell lysis enhancing metal oxide layer, and a cell binding metal oxide layer have been deposited.Type: ApplicationFiled: December 15, 2006Publication date: October 11, 2007Inventors: In-ho Lee, Jun-hong Min, Young-rok Kim, Jae-ho You, Chang-eun Yoo, Jeong-gun Lee, Ki-woong Han
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Publication number: 20070141583Abstract: This invention is related to a method for rapidly identifying regions of the genome to which specific proteins bind, or identifying specific proteins bound to a region of the genome in vivo.Type: ApplicationFiled: December 20, 2005Publication date: June 21, 2007Inventors: Weiwei Li, Jessica M. Li
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Patent number: 7214508Abstract: The present invention relates to the coarse clarification of lysed cell material from microorganisms and particularly to a method of obtaining nucleic acids. In certain embodiments, the present invention provides a method of coarsely clarifying cell lysate from microorganisms that comprises: lysing the microorganisms under the normal atmospheric pressure to form a flocculent precipitate in the lysate, compressing the flocculent precipitate in the lysate by reducing or increasing the pressure of the atmosphere surrounding the lysate in relation to the normal atmospheric pressure to form a compressed flocculent phase and a liquid phase, and separating the two phases.Type: GrantFiled: February 20, 2003Date of Patent: May 8, 2007Assignee: Qiagen GmbHInventors: Joerg Hucklenbroich, Markus Mueller
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Patent number: 7125673Abstract: The present invention provides a method, referred to as CpG retrieval, to overcome incomplete bisulfite modification of DNA recovered from formalin-fixed tissue samples. The method involves boiling deparaffinized tissue samples in citrate buffer, followed by DNA extraction for promoter methylation analysis. In general, the extracted DNA is further modified by sodium bisulfite and then subjected to a method of promoter methylation analysis. The present invention also reports that hypermethylation of ataxia-telangiectasia-mutated gene promoter is correlated with decreased overall survival in patient with head and neck squamous cell carcinoma.Type: GrantFiled: September 23, 2004Date of Patent: October 24, 2006Inventors: Chun-Yang Fan, Thomas Brent
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Patent number: 7122304Abstract: The invention relates to storage of nucleic acid (particularly mRNA) on a solid support and to using such nucleic acid in nucleic acid synthesis or amplification reactions. In a preferred aspect, the invention provides synthesis of cDNA and cDNA libraries.Type: GrantFiled: November 30, 2000Date of Patent: October 17, 2006Assignee: Whatman, Inc.Inventors: Mindy D. Goldsborough, Donna K. Fox
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Patent number: 7109322Abstract: A process for the depletion or removal of endotoxins from preparations containing active ingredients designated for therapeutical use which are obtained from natural sources by genetic engineering and/or biotechnology by treatment with chromatographic material wherein said natural source are lysed, the fractions obtained are optionally centrifuged, filtrated or treated with affinity chromatographic methods; said fractions are preincubated with an aqueous salt solution and detergents, treated with anion exchange material and then washed with another salt solution, and the active ingredients are eluted from the anion exchanger, followed by further purification in a per se known manner.Type: GrantFiled: September 26, 2002Date of Patent: September 19, 2006Assignee: Qiagen GmbHInventors: Metin Colpan, Peter Moritz, Joachim Schorr
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Patent number: 7078224Abstract: Methods are disclosed for using paramagnetic particles to concentrate or harvest cells. Methods are also disclosed for clearing a solution of disrupted biological material, such as a lysate of cells or a homogenate of mammalian tissue. Methods are also disclosed for using paramagnetic particles to isolate target nucleic acids, such as RNA or DNA, from a solution cleared of disrupted biological material using the same type or a different type of paramagnetic particle. Kits are also disclosed for use with the various methods of the present invention. Nucleic acids isolated according to the present methods and using the present kits are suitable for immediate use in downstream processing, without further purification.Type: GrantFiled: December 30, 1999Date of Patent: July 18, 2006Assignee: Promega CorporationInventors: Rex M. Bitner, Craig E. Smith, Douglas H. White, Braeden L. Butler, Jacqui Sankbeil
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Patent number: 7063975Abstract: A power semiconductor device includes a substrate having an upper surface and a lower surface. The substrate has a trench. First and second doped regions are provided proximate the upper surface of the substrate. A first source region is provided within the first doped region. A second source region is provided within the second doped region. A gate is provided between the first and second source regions. The gate includes a first portion extending downward into the trench. A depth of the trench is no more than a depth of the first doped region.Type: GrantFiled: October 3, 2003Date of Patent: June 20, 2006Assignee: IXYS CorporationInventors: Vladimir Tsukanov, Nathan Zommer
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Patent number: 7011943Abstract: The present invention provides a method and apparatus for to conduct transgenic and targeted mutagenesis screening of genomic DNA. This invention also provides a system for screening DNA for a designated genetic sequence. The system includes a computer having a processor, memory and web browser, wherein the computer receives instructions concerning the designated genetic sequence and other screening parameter selection from a remote user via a form of electronic communication, and an automatic screening device that analyzes samples of genomic DNA for the designated sequence.Type: GrantFiled: September 4, 2001Date of Patent: March 14, 2006Assignee: Transnetyx, Inc.Inventor: Timothy A. Hodge
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Patent number: 6989249Abstract: A method for the indirect extraction of DNA greater than 300 kb in size from non-cultivatable organisms contained in an environmental sample is disclosed. The method comprises isolating the organisms from the sample and embedding the isolated organisms in a block of agarose where the organisms are subsequently lysed and the DNA subjected to a first alternating field electrophoresis to extract DNA fragments which are large in size and separate them from the cell debris. The first electrophoretic migration is followed by an enzymatic restriction step and additional electrophoretic migrations. The invention also encompasses DNA obtained by the method.Type: GrantFiled: October 28, 2002Date of Patent: January 24, 2006Assignee: LibragenInventors: Renaud Nalin, Patrick Robe, Van Tran Van
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Patent number: 6977178Abstract: The present invention provides a method and apparatus for to conduct transgenic and targeted mutagenesis screening of genomic DNA. This invention also provides a system for screening DNA for a designated genetic sequence. The system includes a computer having a processor, memory and web browser, wherein the computer receives instructions concerning the designated genetic sequence and other screening parameter selection from a remote user via a form of electronic communication, and an automatic screening device that analyzes samples of genomic DNA for the designated sequence.Type: GrantFiled: September 3, 2002Date of Patent: December 20, 2005Assignee: TransnetYX, Inc.Inventors: Timothy A. Hodge, Shawn O'Malley
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Patent number: 6969603Abstract: Method for isolating DNA contained in a biological sample. The method includes combining in a solution a DNA-containing biological sample, a salt, a cationic surfactant, and a DNA-binding carrier, the solution having a salt concentration higher than the DNA precipitation inhibition-initiating concentration, to lyse the DNA-containing biological sample and to bind DNA to the DNA-binding carrier while in the solution to form a bound DNA-carrier. The method also includes separating the DNA-bound carrier from other components. The method further includes dissociating the bound DNA from the DNA-binding carrier. The method still further includes recovering dissociated DNA.Type: GrantFiled: October 31, 2001Date of Patent: November 29, 2005Assignees: Riken, Kabushiki Kaisha DnaformInventors: Yoshihide Hayashizaki, Piero Carninci
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Patent number: 6933379Abstract: The invention relates to a method for linking nucleic acid and/or glycosaminoglycan or glycosaminoglycan mimetics to a polar/hydrophilic material, characterized by contacting a nucleic acid and/or glycosaminoglycan and a polar/hydrophilic material with each other in the presence of a solution being 20 to 100 percent saturated with a non-chaotropic salt and removing said solution from the nucleic acid and/or glycosaminoglycan—polar/hydrophilic material.Type: GrantFiled: March 30, 2001Date of Patent: August 23, 2005Assignee: Angiomed GmbH & Co. Medizintechnik KGInventors: Antonius H. M. S. M. Van Kuppevelt, Jacobus Henricus Veerkamp, Thiemo Arnim Blank
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Patent number: 6919200Abstract: A purification apparatus, kit and method for purifying DNA, RNA, proteins, antigens, antibodies and cells. The apparatus has a wand and a reservoir tube. The wand is made of a cap, a sample collection assembly and an elongated shaft connecting the cap to the sample collection assembly. The sample collection assembly has a series of microstructures on its surface, or microparticles enclosed within it for increasing the surface area of the sample collection assembly. The increased surface area permits maximum exposure to and binding of target molecules thereto. The reservoir tube associated with the wand has one end defining an opening and a second end that is closed and preferably cone or cylindrical shaped. The cap of the wand securely and sealingly fastens to the open end of the reservoir tube with the shaft and the sample collection assembly fitting easily inside the reservoir tube.Type: GrantFiled: November 22, 1999Date of Patent: July 19, 2005Assignee: The United States of America as represented by the Secretary of the ArmyInventor: Sofi M. Ibrahim
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Patent number: 6838238Abstract: This invention provides a method for identifying one or more complexes from a library of complexes, wherein said complex or complexes are selected for their ability to perform a preselected or desired function on a target molecule or by having a pre-selected structure, each complex being designated a morphatide, said method comprising: (a) preparing a library of morphatides, comprised of: (i) a scaffolding component selected from the group consisting of nucleic acid, nucleic acid like molecule or nucleic acid analog having one or more regions of randomized sequence; (ii) one or more linker components; and (iii) one or more agent molecules or type of agent molecules, linked to the scaffolding component by one or more type of linker components; and (b)screening the library of morphatides prepared in step (a) by contacting, binding, or associating the morphatides with one or more suitable target molecules upon which a morphatide performs a preselected or desired function or to which a morphatide binds or associaType: GrantFiled: April 14, 1997Date of Patent: January 4, 2005Assignee: Invitrogen CorporationInventor: Jay M. Short
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Publication number: 20040265785Abstract: In order to achieve a reproducible treatment for materials of biological origin such as membranes, skins, vessels, heart valves, tendons and ligaments and, in particular, collagen materials, hydrophobic accompanying substances are, first of all, chemically removed without the accompanying substances being chemically altered. Non-hydrophobic accompanying substances are subsequently removed. The materials treated in this manner are used to form membranes, heart valves or vessels or are further processed to form solutions, fibers, fiber weaves, fibrous tissues or sponges and are optionally populated with cells and/or used for carrying out controlled tissue regeneration.Type: ApplicationFiled: May 24, 2004Publication date: December 30, 2004Applicant: MATRICEL GMBHInventors: Leon Olde Damink, Ingo Heschel
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Patent number: 6833257Abstract: A method for detecting the presence of a target nucleic acid sequence in a sample, said method comprising: (a) adding to a sample suspected of containing said target nucleic acid sequence, a probe specific for said target sequence and DNA duplex binding agent, said probe comprising a reactive molecule able to absorb fluorescence from or donate fluorescent energy to said DNA duplex binding agent, (b) subjecting the thus formed mixture to an amplification reaction in which target nucleic acid is amplified, (c) subjecting said sample to conditions under which the said probe hybridizes to the target sequence, and (d) monitoring fluorescence from said sample. This method can be used for example to monitor amplification reactions such as PCR reactions, such that the amount of target sequence present in the sample may be determined.Type: GrantFiled: May 25, 2000Date of Patent: December 21, 2004Assignee: The Secretary of State for DefenceInventors: Martin A Lee, Roderick Fuerst
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Publication number: 20040241656Abstract: A high-performance bio-tube for purifying DNA and a method thereof are provided. The high-performance bio-tube is provided with bio-particles immobilized therein. The bio-particles have particle sizes not more than 100 &mgr;m and is capable of binding DNA. A biological sample is added in the bio-tube and then a lysis buffer is added to form an admixture of the biological sample and the lysis buffer for lysing the cells to release DNAs. The admixture is maintained in the bio-tube in a period of time sufficiently to make the released DNAs binding to the bio-particles immobilized in the bio-tube. The remaining admixture in the bio-tube is directly discarded. Then, eluting DNAs bound to the bio-particles by feeding an elution solution in the bio-tube.Type: ApplicationFiled: May 29, 2003Publication date: December 2, 2004Inventors: Ming-Shiung Jan, Chin-Horng Wang
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Publication number: 20040241769Abstract: A novel reagent system for use with automated and semi-automated hematology analyzers including an essentially isotonic blood diluting reagent, a blood cell lysing and hemoglobin conversion reagent, and a second lysing reagent for differentiating white blood cells into classes by size and functional characteristics. The diluent reagent enhances properties for counting and sizing blood specimens, while stabilizing cellular volume and cellular integrity for many hours. The blood cell lysing reagent removes red blood cells and enables subsequent enumeration of white blood cells and simultaneous determination of hemoglobin without use of the toxic cyanide anion. The third lysing reagent and a companion quenching differentiates blood cells into classes by size and functional characteristics, based on d.c. impedance volume, conductivity/opacity and light scatter measurements. The companion quenching reagent adjusts pH and conductivity of the final measurement solution to match the analyzer system requirements.Type: ApplicationFiled: March 2, 2004Publication date: December 2, 2004Inventors: Harold Richardson Crews, James Harrison Carter, Michael Norman Elliot
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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: 20040229344Abstract: Environmental samples typically include impurities that interfere with PCR amplification and DNA quantitation. Samples of soil, river water, and aerosol were taken from the environment and added to an aqueous buffer (with or without detergent). Cells from the sample are lysed, releasing their DNA into the buffer. After removing insoluble cell components, the remaining soluble DNA-containing extract is treated with N-phenacylthiazolium bromide, which causes rapid precipitation of impurities. Centrifugation provides a supernatant that can be used or diluted for PCR amplification of DNA, or further purified. The method may provide a DNA-containing extract sufficiently pure for PCR amplification within 5-10 minutes.Type: ApplicationFiled: May 15, 2003Publication date: November 18, 2004Inventors: John M. Dunbar, Cheryl R. Kuske
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Publication number: 20040224344Abstract: This invention presents a set of methods to extract DNA, RNA and protein simultaneously from biomaterials by reagents with high pH. DNA and RNA can be extracted from a upper aqueous phase simultaneously either together as a DNA and RNA mixture or separated DNA and RNA. Protein can be extracted from lower organic phase. The DNA and RNA mixture can be used either as DNA or RNA directly depending on applications without further separation, or as resource for the separated DNA and RNA that can be selected from the DNA and RNA mixture by selective precipitation and/or by selective enzyme digestions. This invention provides the choice of extraction either of DNA and RNA mixture or of separated DNA and RNA simultaneously, as well as extraction of protein from the same piece of biomaterials in high quality, which is very critical for biomaterials with limited resource, such as clinical specimens.Type: ApplicationFiled: May 5, 2004Publication date: November 11, 2004Applicant: BioChain Inc.Inventors: Xiaoliang Han, Zhongdong Liu, Jinming Xia
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Publication number: 20040219534Abstract: A method of extracting RNA from biological systems (cells, cell fragments, organelles, tissues, organs, or organisms) is presented in which a solution containing RNA is contacted with a substrate to which RNA can bind. RNA is withdrawn from the substrate by applying negative pressure. No centrifugation step exceeds thirty seconds. Preferably, the RNA is diluted prior to filtration and one or more wash steps can be used to remove interferents. In one such method, except for DNA shearing and drying steps, centrifugation is not undertaken during the extraction step.Type: ApplicationFiled: May 1, 2003Publication date: November 4, 2004Inventors: Robert Belly, Jacqueline Toner, John Backus
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Publication number: 20040209273Abstract: The present invention relates to a method to isolate plant genes or gene fragments that are regulated by stress, preferably oxidative stress. The method includes isolation of plant material, adaptation of the plant material to stress, differential expression of genes or gene fragments in adapted and nonadapted plant material, and isolation of the differentially expressed genes or gene fragments. The invention further relates to the genes or gene fragments that can be obtained by this method and to the use of these genes or gene fragments to modulate plant stress tolerance.Type: ApplicationFiled: August 25, 2003Publication date: October 21, 2004Inventors: Eva Vranova, Dirk Inze, Frank Van Breusegem, Wim Van Camp
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Publication number: 20040197780Abstract: Described herein is a method in which genomic nucleic acid of a cell can be separated from nucleic acid having a molecular weight that is lower than the molecular weight of the genomic nucleic acid (e.g., plasmid DNA) of the cell directly from a cell growth culture. Also described herein, a method in which genomic nucleic acid can be separated from nucleic acid having a molecular weight that is lower than the molecular weight of the genomic nucleic acid in a cell lysate without the need to prepare a cleared lysate.Type: ApplicationFiled: April 2, 2003Publication date: October 7, 2004Applicant: Agencourt Bioscience CorporationInventors: Kevin McKernan, Junaid Ziauddin
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Publication number: 20040185449Abstract: Methods for preparing samples for use in assays are described. The methods use polyethylene glycol and magnetic particles and are conducted with minimal salt concentrations. The methods are useful for preparing samples such as blood samples for use in hybridization assays such as bDNA assays, immunoassays and PCR. Optimal process conditions that allow for highly sensitive assays are also described.Type: ApplicationFiled: March 20, 2003Publication date: September 23, 2004Inventors: John J. Quinn, Uri Piran, Laurie A. Livshin, Claudine Z. Yannoni
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Publication number: 20040171048Abstract: A minimally invasive transdermal nucleic acid sampling method comprises piercing through the outermost layer of the skin and into the underlying epidermis with a plurality of microprojections. Living skin cells in the underlying epidermis are disrupted, causing them to release their contents including their nucleic acids (i.e., DNA, RNA, fragments thereof or other polynucleic acid material found in the nucleii and/or mitochondria of cells). The nucleic acid is collected on the surfaces of the microprojections and/or in a separate nucleic acid collection reservoir. The collected nucleic acid is then analyzed using standard polymerase chain reaction (PCR) techniques. Optionally a suction device applies a partial vacuum through openings in the microprojection member to the microcuts in the skin for enhanced efflux of intracellular and extracellular (i.e., body) fluids containing the nucleic acid.Type: ApplicationFiled: December 23, 2003Publication date: September 2, 2004Inventors: James A. Matriano, Michel J.N. Cormier
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Patent number: 6780632Abstract: The invention relates to host cells and methods of preparing a substantially RNA-free cellular component, comprising culturing cells producing the cellular component in a medium and lysing the cells to produce a cell lysate, wherein the cell lysate contains the cellular component and sufficient RNase activity to degrade substantially all of the RNA molecules present in the cell lysate. The invention also relates to substantially RNA-free cellular components.Type: GrantFiled: April 14, 1999Date of Patent: August 24, 2004Assignee: Cobra Biologics LimitedInventors: Julian Alexis John Hanak, Steven Geraint Williams