Acellular Exponential Or Geometric Amplification (e.g., Pcr, Etc.) Patents (Class 435/91.2)
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Publication number: 20140308673Abstract: A method for replicating and amplifying a target nucleic acid sequence is described. A method of the invention involves the formation of a recombination intermediate without the prior denaturing of a nucleic acid duplex through the use of a recombination factor. The recombination intermediate is treated with a high fidelity polymerase to permit the replication and amplification of the target nucleic acid sequence. In preferred embodiments, the polymerase comprises a polymerase holoenzyme. In further preferred embodiments, the recombination factor is bacteriophage T4 UvsX protein or homologs from other species, and the polymerase holoenzyme comprises a polymerase enzyme, a clamp protein and a clamp loader protein, derived from viral, bacteriophage, prokaryotic, archaebacterial, or eukaryotic systems.Type: ApplicationFiled: June 24, 2014Publication date: October 16, 2014Inventors: Stephen J BENKOVIC, Frank Salinas
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Publication number: 20140308708Abstract: An enzymatic method for removing sequence errors in nucleic acid molecules are described. The method utilizes a CEL endonuclease that cuts heteroduplexes at mismatch sites containing the errors and an overlap extension polymerase chain reaction to re-assemble the cleaved fragments into full-length nucleic acid molecules free of the errors.Type: ApplicationFiled: April 16, 2013Publication date: October 16, 2014Inventor: Jingdong TIAN
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Publication number: 20140308710Abstract: The present invention discloses methods for assembling a nucleic acid molecule from a set of overlapping oligonucleotides. The method involves contacting a set of overlapping oligonucleotides with a DNA polymerase, a mixture of dNTPs, and a crowding agent to form an assembly mixture. In one embodiment the crowding agent is polyethylene glycol (PEG). The presence of the crowding agent facilitates the nucleic acid assembly process of the invention. The assembly mixture is then subjected to multiple cycles, each cycle comprising an annealing phase, an extension phase, and a denaturation phase, and the desired nucleic acid molecule is thereby assembled. In some embodiments one or more of the phases are time varied.Type: ApplicationFiled: December 11, 2013Publication date: October 16, 2014Applicant: Synthetic Genomics, Inc.Inventors: Zhiqing Qi, Jun Urano, Nicky C. Caiazza, Daniel G. Gibson
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Publication number: 20140308709Abstract: Provided is a polynucleotide including, from the 3? terminus of the polynucleotide to the 5? terminus of the polynucleotide, a first region including a nucleotide sequence complementary to a nucleotide sequence of a portion of a target nucleic acid; a second region including a nucleotide sequence identical to a nucleotide sequence of a portion of the target nucleic acid; and a third region including a nucleotide sequence that self-hybridizes to form a stem-loop structure, and compositions, kits, and methods related thereto.Type: ApplicationFiled: October 10, 2013Publication date: October 16, 2014Inventors: Sea-hee KIM, Joo-won Rhee, Ko-bong Choi
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Patent number: 8859238Abstract: The present invention provides a method that combines the construction of double-stranded target amplification products with one or two single-stranded overhangs with improved production of such target amplification products. The single-stranded overhang(s) can be used for post-amplification capture and subsequent detection/manipulation. The single-stranded overhang(s) enable capture/detection/manipulation without interference from the complementary strand in the double-stranded target amplification product.Type: GrantFiled: May 6, 2011Date of Patent: October 14, 2014Assignee: Quantibact A/SInventors: Uffe Vest Schneider, Gorm Lisby
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Patent number: 8859204Abstract: A method comprises loading a sample portion into a sample chamber which comprises means for minimizing diffusion of the sample portion, subjecting the sample portion to an amplification step, and determining whether the sample portion contains at least one molecule of a target nucleic acid. If the sample portion contains a single molecule of the target nucleic acid, the sample portion would attain a detectable concentration of the target nucleic acid after a single round of amplification. Also, a microfluidic device comprising a sample portion and a sample chamber comprising means for minimizing diffusion of the sample portion. Also, a microfluidic device comprising a sample chamber and an amplification targeting reagent positioned in the first sample chamber.Type: GrantFiled: August 13, 2007Date of Patent: October 14, 2014Assignees: Applied Biosystems, LLC, The United States of America, As Represented by the Secretary, Department of Health and Human ServicesInventors: James F. Brown, Jonathan E. Silver
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Patent number: 8859239Abstract: Next generation sequencing technologies are becoming a preferred method for sequencing nucleic acids and profiling miRNAs. Experimental results disclosed herein show that the most common platform for preparing nucleic acids such as miRNAs for sequencing introduces serious biases. Provided herein are compositions and methods for improved sequencing and miRNA profiling using a set of customized ligation adaptors.Type: GrantFiled: May 11, 2012Date of Patent: October 14, 2014Assignee: City of HopeInventors: Yun Yen, Guihua Sun
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Patent number: 8859199Abstract: The present invention relates to a method of ensuring the effectiveness of the extraction workup from a biological sample of nucleic acid. The inventive method is able to distinguish between possible defects in the extraction of nucleic acid from a sample and possible defects in a subsequent amplification step. The present invention also relates to a packaged array for extracting nucleic acid from a biological sample.Type: GrantFiled: April 4, 2008Date of Patent: October 14, 2014Assignee: Becton, Dickinson and CompanyInventors: Tobin Hellyer, Thomas Fort, Ray A. McMillian
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Publication number: 20140302562Abstract: Provided herein is a microplate for polymerase chain reaction (PCR), comprising a substrate formed of a material that is susceptible to heating PCR samples upon the application of an electromagnetic field and/or electromagnetic energy to said substrate. The substrate provides a PCR ramp rate of at least 5° C./second upon the application of an electromagnetic field and/or electromagnetic energy to said substrate.Type: ApplicationFiled: March 12, 2014Publication date: October 9, 2014Applicant: BJS IP Ltd.Inventor: Nicholas BURROUGHS
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Publication number: 20140302504Abstract: Methods and compositions for the amplification of nucleic acids are disclosed. Amplification methods provided herein may be performed under isothermal conditions. Methods and compositions may include reagents such as nucleic acid polymerases, ligases, and primers.Type: ApplicationFiled: March 15, 2014Publication date: October 9, 2014Inventors: Kamila Belhocine, Josephine Lee, Pranav Patel, Aaron Richardson, Scott Tabakman
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Publication number: 20140302559Abstract: In the method as reported herein the isolation of nucleic acid segments encoding antibody variable domains and the insertion of the isolated nucleic acid segments in eukaryotic expression plasmids is performed without the intermediate isolation and analysis of clonal intermediate plasmids. Thus, in the method as reported herein the intermediate cloning, isolation and analysis of intermediate plasmids is not required, e.g. by analysis of isolated transformed E. coli cells.Type: ApplicationFiled: June 20, 2014Publication date: October 9, 2014Applicant: HOFFMANN-LA ROCHE INC.Inventors: Simone Hoege, Erhard Kopetzki, Dominique Ostler, Stefan Seeber, Georg Tiefenthaler
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Patent number: 8852920Abstract: The present invention relates to a micro-chamber plate and a manufacturing method of the same, more precisely a micro-chamber plate facilitating real-time measurement and analysis of fluorescence obtained from the reaction of multiple reaction solutions containing primers or probes selectively binding to each corresponding gene without cross-contamination in order to analyze biological samples containing numbers of genes and a manufacturing method of the same.Type: GrantFiled: September 22, 2008Date of Patent: October 7, 2014Assignee: Bioneer CorporationInventors: Han Oh Park, Gu-Young Song
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Patent number: 8852892Abstract: A real-time system for determining the geographic movements of an individual or object by sampling particulates contained thereon. The system includes particle collection, sample preparations, and sample analysis using three primary modes of detecting certain particulates. The first mode involves the imaging of pollen, spores, or other biological material which are visible through a light microscope when properly stained or prepared. The second mode involves the use of real-time polymerase chain reaction to amplify and detect target nucleic acid sequences. The third mode involves the use of X-ray diffraction to identify mineral particles. The results from any mode, or any combination of modes, are analyzed by comparison to a reference database containing geographic information and the results are compiled by a controller for visual display.Type: GrantFiled: February 5, 2010Date of Patent: October 7, 2014Assignee: Syracuse UniversityInventors: Donald F. Shepard, David B. Knaebel, D. Anthony Gray
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Patent number: 8852893Abstract: The invention generally relates to methods for detecting a target nucleic acid and a target protein in a single assay.Type: GrantFiled: June 5, 2012Date of Patent: October 7, 2014Assignee: Physicians Choice Laboratory Services, LLCInventor: Anthony P. Shuber
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Patent number: 8852867Abstract: The invention provides methods for amplification of polynucleotide sequences using primers containing single-stranded RNA. The methods employ use of an enzyme capable of cleaving single-stranded RNA, such as RNase I, to degrade a first RNA-containing primer prior to addition of a second RNA-containing primer. The invention also provides compositions and kits for practicing the amplification methods, as well as methods which use the amplification products.Type: GrantFiled: May 9, 2011Date of Patent: October 7, 2014Assignee: Nugen Technologies, Inc.Inventors: Nurith Kurn, Shenglong Wang
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Patent number: 8852863Abstract: The present invention relates to a method for amplifying and detecting nucleic acid sequences in a reaction cartridge comprising, (i) providing a sample comprising at least one nucleic acid molecule, (ii) in a first reaction chamber of the cartridge providing reagents for an amplification reaction, (iii) mixing the sample with the amplification reagents, (iv) amplifying the at least one nucleic acid in the first reaction chamber of the cartridge, (v) transferring at least parts of the amplification reaction into a second and third reaction chamber of the cartridge each comprising a probe set and performing a melting point analysis in order to determine which of the probes has specifically bound a nucleic acid.Type: GrantFiled: May 4, 2010Date of Patent: October 7, 2014Assignee: Qiagen GmbHInventors: Thomas Rothmann, Holger Engel, Ralf Himmelreich, Andy Wende, Rainer Dahlke
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Patent number: 8852894Abstract: A method of detecting an analyte is provided. The method includes providing a sample, a container 110 with a wall 115, and a catalyst for a luminescent reaction. The wall includes a colored portion 115b. The method further comprises forming a reaction in the container and detecting the presence or absence of light emitted from the reaction mixture in the container. Detecting light emitted from the container can comprise detecting light passing through the colored portion. The colored portion can be detected visually and the color can be associated with the identity of an analyte—specific reagent disposed in the container. Kits comprising the container and a catalyst for a luminescent reaction are also provided.Type: GrantFiled: April 19, 2012Date of Patent: October 7, 2014Assignee: 3M Innovative Properties CompanyInventors: Neil Percy, Gregory W. Sitton
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Patent number: 8852866Abstract: Methods for producing and identifying fragments of proteins, and more particularly to methods for generating and identifying soluble protein domains are disclosed based on a method for generating a library of nucleic acid fragments from nucleic acid encoding a desired polypeptide, and more especially a library of essentially, randomly sampled fragments of coding DNA sequence predominantly of defined size range and a method for selecting cloned gene fragments from the library that encode soluble protein domains.Type: GrantFiled: November 8, 2002Date of Patent: October 7, 2014Assignee: Domainex LimitedInventors: Mark McAlister, Renos Savva, Laurence Pearl, Chrisostomos Prodromou, Paul C Driscoll
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Publication number: 20140295439Abstract: Methods and compositions for the amplification of nucleic acids are disclosed. Amplification methods provided herein may be performed under isothermal conditions. Methods and compositions may include reagents such as restriction enzymes, polymerases, ligases, primers, and polynucleotide adaptors.Type: ApplicationFiled: March 15, 2014Publication date: October 2, 2014Inventor: Pranav Patel
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Publication number: 20140296082Abstract: Compositions and methods are described to modify Family B DNA polymerases that contain residual exonuclease activity that interferes with sequencing techniques and with detection of single nucleotide polymorphisms. The compositions are mutant proteins with reduced exonuclease activity compared with presently available “exo?” polymerases, and a sensitive screening assay that enables an assessment of exonuclease activity of any synthetic DNA polymerase.Type: ApplicationFiled: April 8, 2014Publication date: October 2, 2014Applicant: NEW ENGLAND BIOLABS, INC.Inventor: Andrew Gardner
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Publication number: 20140295498Abstract: Methods for amplifying a desired target region of a nucleic acid through rolling circle amplification with a strand-displacing polymerase are provided. Concatameric hairpin products are resolved with endonuclease digestion, and the resulting amplified product hairpins or fragments can be circularized and employed as templates in a subsequent round of amplification. The methods are effective for targeted amplification of even highly repetitive sequences. Compositions, kits, and systems related to or useful in the methods are also described.Type: ApplicationFiled: March 13, 2014Publication date: October 2, 2014Applicants: The Regents of the University of California, Pacific Biosciences of California, Inc.Inventors: Stephen W. Turner, Thang Pham, Paul Hagerman
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Publication number: 20140295500Abstract: Disclosed are DNA polymerases having increased reverse transcriptase efficiency relative to a corresponding, unmodified polymerase. The polymerases are useful in a variety of disclosed primer extension methods. Also disclosed are related compositions, including recombinant nucleic acids, vectors, and host cells, which are useful, e.g., for production of the DNA polymerases.Type: ApplicationFiled: May 20, 2014Publication date: October 2, 2014Inventors: Keith Bauer, Thomas W. Myers, Shawn Suko
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Publication number: 20140295499Abstract: Disclosed are mutant DNA polymerases having increased 3?-mismatch discrimination relative to a corresponding, unmodified polymerase. The mutant polymerases are useful in a variety of disclosed primer extension methods. Also disclosed are related compositions, including recombinant nucleic acids, vectors, and host cells, which are useful, e.g., for production of the mutant DNA polymerases.Type: ApplicationFiled: March 31, 2014Publication date: October 2, 2014Applicant: Roche Molecular Systems, Inc.Inventors: Fred Reichert, Keith Bauer, Thomas W. Myers
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Publication number: 20140295440Abstract: Methods and compositions for the amplification of nucleic acids and generation of concatemers are disclosed. Amplification methods provided herein may be performed under isothermal conditions. Methods and compositions may include reagents such nucleic acid polymerases and primers.Type: ApplicationFiled: March 15, 2014Publication date: October 2, 2014Inventors: Kamila Belhocine, Josephine Lee, Pranav Patel, Aaron Richardson, Scott Tabakman
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Patent number: 8846317Abstract: The present invention provides methods and systems for real-time measurements of PCR with multiplexing capability. Certain embodiments relate to methods and systems that use fluorescently encoded superparamagnetic microspheres for the immobilization of amplification products during the PCR process, and an imaging chamber of a measurement device that is also capable of controllable thermal cycling for assisting the PCR process.Type: GrantFiled: September 18, 2012Date of Patent: September 30, 2014Assignee: Luminex CorporationInventors: Douglas F. Whitman, Charles J. Collins
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Patent number: 8846315Abstract: Provided herein are genetic variants associated with development of a condition of interest (e.g., Alzheimer's disease). Methods of treatment with an active agent (e.g., with a particular active agent and/or at an earlier age) is also provided, upon detecting a genetic variant described herein. In some embodiments, the genetic variant is a deletion/insertion polymorphism (DIP) of the TOMM40 gene.Type: GrantFiled: February 17, 2011Date of Patent: September 30, 2014Assignee: Zinfandel Pharmaceuticals, Inc.Inventor: Allen D. Roses
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Patent number: 8846316Abstract: The disclosure relates to a method for determining incidence of liver cancer in a subject, including detecting methylation level or expression level of one microRNA miR-129-2 in a bio-sample from the subject. In the case that the methylation level of the microRNA in the bio-sample is higher or the expression level of the microRNA in the bio-sample is lower relative to that of the corresponding microRNA in a control sample, indicates that the subject is predisposed to or afflicted with liver cancer.Type: GrantFiled: November 26, 2012Date of Patent: September 30, 2014Assignee: Industrial Technology Research InstituteInventors: Chang-Yi Lu, Meng-Tsung Tien, Cheng-Tao Wu, Yih-Huei Uen, Kai-Yuan Lin
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Patent number: 8846349Abstract: This invention relates to a rapid method for detection and characterization of Escherichia coli bacteria serotype O157:H7 based on the presence of nucleic acid sequences, in particular, to a PCR-based method for detection, and to oligonucleotide molecules and reagents and kits useful therefore. This method is preferably employed to detect E. coli O157:H7 in a food or water sample, such as a beef enrichment. The present invention further relates to replication compositions and kits for carrying out the method of the present invention.Type: GrantFiled: July 20, 2010Date of Patent: September 30, 2014Assignee: E.I. du Pont de Nemours and CompanyInventor: Frank R. Burns
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Patent number: 8846341Abstract: The present invention relates to the discovery that of a panel of serum or plasma markers may be used to diagnose Idiopathic Pulmonary Fibrosis (“IPF”) and distinguish this condition from other lung ailments. It further relates to the identification of markers associated with IPF disease progression.Type: GrantFiled: March 3, 2011Date of Patent: September 30, 2014Assignees: University of Pittsburgh—of the Commonwealth System of Higher Education, The University of Chicago, Instituto Nacional de Enfermedades Respiratorias Ismael Cosio Villegas—D.F.Inventors: Naftali Kaminski, Kevin F. Gibson, Bernadette R. Gochuico, Thomas J. Richards, Ivan Rosas, Kazuhisa Konishi, Moises Selman, Jose David Herazo-Maya, Imre Noth
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Patent number: 8846347Abstract: The invention provides methods for preparing DNA sequencing libraries by assembling short read sequencing data into longer contiguous sequences for genome assembly, full length cDNA sequencing, metagenomics, and the analysis of repetitive sequences of assembled genomes.Type: GrantFiled: February 5, 2013Date of Patent: September 30, 2014Assignee: University of WashingtonInventors: Jay Shendure, Joseph Hiatt, Rupali Patwardhan, Emily Turner
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Patent number: 8846312Abstract: The present invention provides rapid and accurate methods, primers, probes and kits for identifying the presence of a certain flaviviruses in a sample. Flaviviruses that can be detected include members of the Japanese encephalitis virus serogroup, Dengue virus, St. Louis encephalitis virus, Montana myotis leukoencephalitis virus, Modoc virus, and Yellow Fever virus. The primers and probes of the invention can hybridize to regions in the 3? untranslated region of the viral genomes to be detected.Type: GrantFiled: April 16, 2013Date of Patent: September 30, 2014Assignee: Roche Molecular Systems, Inc.Inventor: Karen K. Y. Young
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Patent number: 8846313Abstract: A device comprising a rigid substrate, a flexible cover element at least partially covering the substrate, a first structure formed in the substrate, adapted for accommodating liquids and adapted for releasing contents of one or more cells, spores, or viruses, the contents including the target molecules, a second structure formed in the substrate, adapted for accommodating liquids and comprising at least one binding member adapted for capturing the target molecules and for determining a value indicative of the presence and/or amount of the target molecules, a micro fluidic network interconnecting at least the first structure and the second structure, and an actuator member adapted for effecting a fluid flow between the first structure and the second structure by pressing the flexible cover element against the substrate to selectively close a portion of the micro fluidic network.Type: GrantFiled: November 6, 2007Date of Patent: September 30, 2014Assignee: Clondiag GmbHInventors: Katrin Steinmetzer, Eugen Ermantraut, Torsten Schulz, Thomas Kaiser, Thomas Ullrich
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Patent number: 8846883Abstract: Oligonucleotide chemistry is central to the advancement of core technologies such as DNA sequencing, forensic and genetic analysis and has impacted greatly on the discipline of molecular biology. Oligonucleotides and their analogues are essential tools in these areas. They are often produced by automated solid-phase phosphoramidite synthesis but it is difficult to synthesize long DNA and RNA sequences by this method. Methods are proposed for ligating oligonucleotides together, in particular the use of an azide-alkyne coupling reaction to ligate the backbones of oligonucleotides together to form longer oligonucleotides that can be synthesized using current phosphoramidite synthesis methods.Type: GrantFiled: March 1, 2012Date of Patent: September 30, 2014Assignee: University of SouthhamptonInventors: Tom Brown, Afaf Helmy El-Sagheer
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Patent number: 8840848Abstract: Systems and methods for processing and analyzing samples are disclosed. The system may process samples, such as biological fluids, using assay cartridges which can be processed at different processing locations. In some cases, the system can be used for PCR processing. The different processing locations may include a preparation location where samples can be prepared and an analysis location where samples can be analyzed. To assist with the preparation of samples, the system may also include a number of processing stations which may include processing lanes. During the analysis of samples, in some cases, thermal cycler modules and an appropriate optical detection system can be used to detect the presence or absence of certain nucleic acid sequences in the samples. The system can be used to accurately and rapidly process samples.Type: GrantFiled: January 23, 2013Date of Patent: September 23, 2014Assignee: Beckman Coulter, Inc.Inventor: Charles S. Kraihanzel
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Patent number: 8841095Abstract: The invention generally relates to methods for detecting a target nucleic acid and a target protein in a single assay.Type: GrantFiled: April 23, 2013Date of Patent: September 23, 2014Assignee: Physicians Choice Laboratory Services, LLCInventor: Anthony P. Shuber
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Patent number: 8841075Abstract: A Homologous Pairing Capture Assay is described which enables detection of coalignment between homologous DNA sequences. The assay involves ligating closely positioned homologous sequences to each other thereby generating head-to-head ligation products or inverted repeats. DNA fragments containing an inverted repeat are then converted into hairpin DNA molecules. The hairpin DNA molecules can then be readily separated from DNA molecules free of inverted repeats. Also described are various diagnostic applications and kits relating to the assay.Type: GrantFiled: April 12, 2011Date of Patent: September 23, 2014Assignee: Cleveland State UniversityInventors: G. Valentin Börner, Neeraj Joshi
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Patent number: 8841093Abstract: The invention provides an apparatus that can be used in methods of preparing, amplifying, detecting, and/or optionally selecting for further analysis the genomic material from an organism for the rapid detection and/or classification of an organism in a sample (e.g., screening for, identifying, quantifying, and/or optionally further analyzing, e.g., sequencing, the genomic material of the organism). The invention further provides methods of using the apparatus, e.g., in combination with novel SGP primers for improved use in waveform-profiling methods of DNA amplification. It is an object of the invention to provide an apparatus for fully automated analysis of genomic material, and multiple methods of using the apparatus that are beneficial to society, e.g., the apparatus may be used in methods of screening for, identifying, quantifying, and/or selecting genomic material for further analysis (e.g., sequencing) in relation to monitoring a source for the presence of contaminating organisms.Type: GrantFiled: May 15, 2009Date of Patent: September 23, 2014Assignee: Canon U.S. Life Sciences, Inc.Inventors: Toru Takahashi, Hiroshi Inoue
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Patent number: 8841094Abstract: The present invention provides a method which can achieve the homologous recombination of a gene of interest selectively, and a recombinant DNA molecule produced by the method. A homologous recombination method which uses a PCR product and a linearized vector is disclosed. The PCR product comprises a sequence for a target gene and amplification primer sequences P1 and P2 on both terminal ends. The vector contains homologous recombination regions VP1 and VP2 which respectively comprise nucleotide sequences homologous to P1 and P2, and at least one a homologous recombination region VT (VT1 and/or VT2), which comprises a nucleotide sequence homologous to a sequence T (T1 and/or T2), T sequences are sequence parts internal to P1 and/or P2 as well as sequence parts on the terminal side of VP1 and/or VP2 (provided that at least one T sequence has a nucleotide sequence specific to the target gene).Type: GrantFiled: March 6, 2009Date of Patent: September 23, 2014Assignee: National University Corporation University of ToyamaInventors: Nobuyuki Kurosawa, Masaharu Isobe
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Patent number: 8841071Abstract: The invention generally relates to methods for sample multiplexing. In certain embodiments, methods of the invention obtaining a plurality of different reactant molecules, attaching a unique identifier to the reactant molecules, and forming a droplet including the reactant molecules.Type: GrantFiled: May 31, 2012Date of Patent: September 23, 2014Assignee: Raindance Technologies, Inc.Inventor: Darren Link
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Publication number: 20140273090Abstract: An amino acid chain of interest is selected or captured for amplification. The amino acid chain of interest including at least one amino acid sequence is denatured and read. The read amino acid sequence is transcribed to synthesize a mRNA equivalent. The amino acid chain of interest is then amplified using a biological system.Type: ApplicationFiled: March 12, 2014Publication date: September 18, 2014Applicant: Arizona Board of Regents, a body corporate of the State of Arizona, acting for and on behalf ofInventor: Rolf U. Halden
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Publication number: 20140272999Abstract: The present invention relates to a simple and efficient method to isolate and purify nucleic acids, preferably genomic DNA, from complex samples compared with available methods, by using a ligand which relies on hydrogen bonding to purify the nucleic acids. Preferably the ligand is bound to magnetic beads/particles. More closely the method comprises adding a sample comprising nucleic acid to a polymer having neutral charge; reversibly binding said nucleic acid to said polymer by hydrogen bonding under pH conditions <5; washing said polymer; and eluting said nucleic acid from said polymer under conditions of pH >5. The method is very suitable for sample preparation of nucleic acids, for example for PCR applications.Type: ApplicationFiled: October 25, 2012Publication date: September 18, 2014Applicant: GE HEALTHCARE BIO-SCIENCES ABInventors: Camilla Estmer Nilsson, Johan Ohman
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Publication number: 20140273102Abstract: Fusion proteins having thermostable blunt-end ligase activity are provided. Blunt-end ligases are useful for DNA amplification, sequencing, production of recombinant DNA and recombinant fusion proteins, and other purposes. These thermostable blunt-end DNA ligases are useful in ligation schemes which include, e.g., an incubation at about 60-65° C. or higher, or as high as about 94° C., or at other temperatures. The ligases disclosed herein may enable high temperature blunt-end ligation without need for molecular crowding agents, and so may be useful for many nucleic acid ligation-amplification schemes, e.g., ones which operate at a uniform temperature (e.g., at about 60° C. or higher), including ones which require temperature cycling, e.g., from about 94° C. to about 60° C. (or higher) for one, two, three, or more cycles.Type: ApplicationFiled: March 15, 2014Publication date: September 18, 2014Applicant: Theranos, Inc.Inventor: Frederick Christians
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Publication number: 20140273099Abstract: Methods and devices for non-thermal PCR amplification of nucleic acid sequences. An electrical potential is applied to cause non-thermal separation of strands of a double-stranded nucleic acid or double-stranded nucleic acid/primer extension product.Type: ApplicationFiled: March 14, 2013Publication date: September 18, 2014Applicant: THE REGENTS OF THE UNIVERSITY OF CALIFORNIAInventor: THE REGENTS OF THE UNIVERSITY OF CALIFORNIA
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Publication number: 20140273101Abstract: A nucleic acid amplification reaction device includes a rotary body having a mounting portion on which a cartridge, which includes a tube that has a plug containing an eluate in which a nucleic acid having bound to nucleic acid-binding solid-phase carriers is eluted from the carriers and a nucleic acid amplification reaction container that is in communication with the tube and contains oil undergoing phase separation from the eluate, is mounted; and a heater forming a temperature gradient inside the nucleic acid amplification reaction container, in which when the posture of the cartridge is changed due to the rotation of the rotary body, a droplet of the eluate introduced from the tube moves inside the nucleic acid amplification reaction container.Type: ApplicationFiled: March 12, 2014Publication date: September 18, 2014Applicant: SEIKO EPSON CORPORATIONInventors: Hiroshi KOEDA, Fumio TAKAGI
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Publication number: 20140273100Abstract: A cDNA synthesis method includes: mixing a lysis solution containing a chaotropic substance and a nucleic acid-binding solid-phase carrier in a sample containing a ribonucleic acid (RNA), thereby adsorbing the RNA on the carrier; reverse-transcribing the RNA adsorbed on the carrier while keeping the RNA adsorbed on the carrier in a reverse transcription reaction mixture, thereby synthesizing cDNA; and eluting the synthesized cDNA with an eluent.Type: ApplicationFiled: March 11, 2014Publication date: September 18, 2014Applicant: Seiko Epson CorporationInventors: Yuji Saito, Fumio Takagi
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Patent number: 8835116Abstract: Methods are provided for identification of genes that are imprinted. In another embodiment methods are provided for identification and analysis of genes whose expression shows allelic imbalance. The expression products transcribed from genes that are present in the genome as two or more alleles may be distinguished by hybridization to an array designed to interrogate individual alleles. Genes whose transcription products are present in amounts that vary from expected are candidates for allelic imbalance, imprinting and imprinting errors.Type: GrantFiled: November 2, 2012Date of Patent: September 16, 2014Assignee: Affymetrix, Inc.Inventor: Giulia Kennedy
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Patent number: 8835115Abstract: The present disclosure provides methods of identifying subjects having an increased likelihood of developing one or more adverse side effects resulting from administration of a microtubule-stabilizing agent. In particular examples, the method includes determining whether the subject has an ABCB1 predictive polymorphism for microtubule-stabilizing agent-induced toxicity, wherein the presence of such a polymorphism indicates that the subject has an increased risk of developing microtubule-stabilizing agent induced adverse effects. Examples of ABCB1 predictive polymorphisms include 2677G>T/A and 3435C>T. Also provided are methods of modifying microtubule-stabilizing agent therapy in a subject identified as having one or more ABCB1 predictive polymorphisms. Kits and isolated nucleic acid molecules that can be used in the disclosed methods are also provided.Type: GrantFiled: July 13, 2007Date of Patent: September 16, 2014Assignees: The United States of America as represented by the Secretary, Department of Health and Human Services, Universitätsklinikum FreiburgInventors: William D. Figg, Klaus Mross, Dirk Behringer, Alex Sparreboom, Tristan Sissung, Stephan Mielke
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Patent number: 8835111Abstract: The invention relates to methods of evaluating MS severity based on analysis of single nucleotide polymorphisms (SNPs) and to products and kits for use in such methods.Type: GrantFiled: March 12, 2010Date of Patent: September 16, 2014Assignee: Brainco Biopharma S.L.Inventors: David Arteta, Marta Artieda, Diego Tejedor, Antonio Martinez, Laureano Simon, Bart A. Crusius, Salvador Pena, Madeleine Sombekke, Bernard Uitdehaag, Chris Polman
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Publication number: 20140255943Abstract: Provided herein are methods, compositions and kits to extract and relatively enrich by physical separation or amplification short base pair nucleic acid in the presence of a high background of genomic material (e.g., host or maternal nucleic acids).Type: ApplicationFiled: February 14, 2014Publication date: September 11, 2014Applicant: SEQUENOM, INC.Inventors: Carolyn R. Hoyal-Wrightson, Andreas Braun, Karsten E. Schmidt
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Publication number: 20140255925Abstract: The invention provides a provides improvements to assays that employ RNase H cleavage for biological applications related to nucleic acid amplification and detection, where the RNase H has been reversibly inactivated.Type: ApplicationFiled: March 15, 2013Publication date: September 11, 2014Applicant: Integrated DNA TechnologiesInventors: Joseph Alan WALDER, Mark Aaron BEHLKE, Scott D. ROSE, Joseph DOBOSY, Susan Marie RUPP