Abstract: The present invention provides methods for reducing the complexity of a nucleic acid sample to interrogate a collection of target sequences. Complexity reduction can be accomplished by fragmenting the nucleic acid sample with a restriction enzyme that has at least one variable position in the recognition sequence. In some aspects adaptors that ligate to some but not all possible overhangs generated by digestion are ligated to the fragments. This selective adaptor ligation allows for selective amplification of a subset of the fragments using primers complementary to the adaptor sequence. In another aspect primers that are complementary to a subset of the fragments after adaptor ligation are used for amplification. Amplified fragments may be analyzed to genotype polymorphisms by hybridization to an array of probes that are complementary to target sequences that will be amplified.
Abstract: The invention provides the use of tetraethylene glycol dimethyl ether for adsorbing nucleic acids to solid phases such as those with silica surfaces. To this end, the invention also provides compositions comprising TDE. Methods are disclosed and claimed to purify nucleic acids from samples, as well as kits useful for performing these methods. Particularly, the invention encompasses methods for the purification of nucleic acids with low molecular weight. The nucleic acids purified by a method of the invention are suited for assays aiming at the detection of a target nucleic acid.
Type:
Grant
Filed:
June 10, 2009
Date of Patent:
February 28, 2012
Assignee:
Roche Diagnostics Operations, Inc.
Inventors:
Horst Donner, Frank Bergmann, Nina Lassonczyk, Manfred Watzele, Marcus Schmid
Abstract: This invention relates to non-radioactive markers that facilitate the detection and analysis of nascent proteins translated within cellular or cell-free translation systems. Nascent proteins containing these markers can be rapidly and efficiently detected, isolated and analyzed without the handling and disposal problems associated with radioactive reagents. Preferred markers are dipyrrometheneboron difluoride (4,4-difluoro-4-bora-3a,4a-diaza-s-indacene) dyes.
Type:
Grant
Filed:
June 23, 2005
Date of Patent:
February 14, 2012
Assignee:
Ambergen, Inc.
Inventors:
Sadanand Gite, Jennifer A. McCullough, Kenneth J. Rothschild
Abstract: It is intended to provide a means and a method whereby a molecule for DNA detection at an elevated accuracy can be provided, different form the existing DNA detection system with the use of a molecular beacon. A molecular beacon wherein fluorescent organic groups capable of forming an excimer are bonded to the 3? and 5? ends of a single-stranded oligonucleotide to be hybridizable with a subject oligonucleotide and the switching of monomer emission/excimer emission is utilized; and a method of detecting SNP by using this molecular beacon.
Abstract: 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.
Abstract: Methods of recombining nucleic acids, including homologous nucleic acids, are provided. Families of gene shuffling oligonucleotides and their use in recombination procedures, as well as polymerase and ligase mediated recombination methods are also provided.
Type:
Grant
Filed:
November 30, 2007
Date of Patent:
October 4, 2011
Assignee:
Codexis Mayflower Holdings, LLC
Inventors:
Andreas Crameri, Willem P. C. Stemmer, Jeremy Minshull, Steven H. Bass, Mark Welch, Jon E. Ness, Claes Gustafsson, Phillip A. Patten
Abstract: This invention provides a method of gene expression analysis that enables extensive gene expression analysis and simultaneous analysis of multiple samples of organisms for which genomic analysis has not yet been advanced. In this method, tags each comprising an oligonucleotide of more than 25 bp for identifying expressed genes, wherein the 3?-end of the tag is defined by a cleavage site of a type III restriction enzyme and the 5?-end thereof is defined by a cleavage site of another restriction enzyme located closest to the 3?-end of the cDNA of such genes, are immobilized on a solid support, gene-containing samples are hybridized to the solid support, and the signals emitted from the genes hybridized to the tags are detected to analyze the gene expression profiles in the samples.
Abstract: A method for DNA reassembly after random fragmentation, and its application to mutagenesis of nucleic acid sequences by in vitro or in vivo recombination is described. In particular, a method for the production of nucleic acid fragments or polynucleotides encoding mutant proteins is described. The present invention also relates to a method of repeated cycles of mutagenesis, shuffling and selection which allow for the directed molecular evolution in vitro or in vivo of proteins.
Abstract: The invention provides turnover probes for use in a variety of detection assays, for example, nucleic acid detection assays. In addition, the invention provides assays, for example, nucleic acid detection assays, using such turnover probes.
Type:
Grant
Filed:
May 3, 2006
Date of Patent:
June 28, 2011
Assignee:
Ensemble Therapeutics Corporation
Inventors:
James M. Coull, Lawrence A. Haff, Joshua A. Bittker, Xiaoyu Li, Larry W. McLaughlin
Abstract: A hybrid formed of a probe nucleic acid immobilized onto a substrate and a sample nucleic acid, in which the target strand of the sample nucleic acid is longer than the probe nucleic acid, so that at least a 5? end portion of the target strand extends upstream the target sequence; a 3? end of the target strand is the same as a 3? end of the target sequence or extends downstream the target sequence by at least one base from the 3? end of the target sequence; and provided that when the number of bases of the extending 5? portion is designated as L1, and the number of bases of the extending 3? portion of the target strand is designated as L2, a value of L1/L2 falls within the range of 0 to 1.5 both inclusive. This hybrid is very stable so that useful for detecting a small amount of nucleic acid specifically with high sensitivity.
Abstract: The present invention provides a method for determining the base sequence of a DNA. According to the method for determining the base sequence of a DNA of the present invention, a probe is used, which is a probe having a protruding end and identification-labeled according to the species of the base at the protruding end, containing a recognition sequence of a class IIS restriction enzyme, to carry out simultaneously in a chain reaction, for a plurality of DNAs to be analyzed, ligation of the end base of a DNA to be analyzed and a probe and cleavage of the end base of the DNA to be analyzed, allowing the base sequence to be determined sequentially by a single molecule spectrofluorimetry method, such that an effective determination of the base sequence of a DNA becomes possible.
Type:
Grant
Filed:
February 20, 2007
Date of Patent:
May 31, 2011
Assignees:
National University Corporation Hokkaido University, Nikon Corporation
Abstract: Systems and methods for analysis of polymers, e.g., polynucleotides, are provided. The systems are capable of analyzing a polymer at a specified rate. One such analysis system includes a structure having a nanopore aperture and a molecular motor, e.g., a polymerase, adjacent the nanopore aperture.
Type:
Grant
Filed:
November 25, 2009
Date of Patent:
May 24, 2011
Assignees:
President and Fellows of Harvard College, Regents of the University of California, Agilent Technologies, Inc.
Inventors:
Mark Akeson, Daniel Branton, David W. Deamer, Jeffrey R. Sampson
Abstract: A device and kit for pulsing a biological sample with a pulsing agent is disclosed. The biological sample so pulsed is subsequently stabilized and the device or kit provides a control reaction. Applications for the described device and kit are found in the field of medical diagnostics, particularly immunology.
Type:
Grant
Filed:
October 17, 2008
Date of Patent:
May 24, 2011
Assignee:
Universite Libre de Bruxelles
Inventors:
Patrick Stordeur, Michel Goldman, Marius Tuijnder
Abstract: A method of detecting a molecule associated with viability of one or more cells or organisms in a sample comprises the initial step of contacting the sample with an enzyme, which enzyme is capable of adding or removing a chemical moiety to or from a nucleic acid molecule in the presence of the molecule associated with viability of the of the one or more cells or organisms. This thereby generates a novel detectable nucleic acid molecule. The next step involves detecting the presence of the molecule associated with viability of the one or more cells or organisms by detecting the novel nucleic acid molecule generated only in the presence of the molecule associated with viability of the one or more cells or organisms. A most preferred molecule associated with viability is ATP, although NAD may also be detected. A preferred enzyme for use in the methods is ligase.
Abstract: The disclosure is related generally to methods for testing mammary fluid (including milk) to establish or confirm the identity of the donor of the mammary fluid. Such methods are useful in the milk-bank business to improve safety.
Type:
Grant
Filed:
March 20, 2008
Date of Patent:
May 17, 2011
Assignee:
Prolacta Bioscience, Inc.
Inventors:
Elena M. Medo, Martin L. Lee, David J. Rechtman
Abstract: A non-amplification multiple quantitative detection kit is characterized in that there are provided a single strand poly nucleotide having a base sequence designed to recognize a specific gene portion of a multiple pathogenic microorganisms to be detected by a fishing probe, and a double strand poly nucleotide indicated with a tag having a base sequence designed to recognize different gene portions which does not overlap with the recognition portion of the fishing probe in the specific genes of the multiple pathogenic microorganisms to be detected by a reporter probe.
Type:
Grant
Filed:
April 13, 2007
Date of Patent:
May 3, 2011
Assignees:
Mokpo National University Industry-Academic Cooperation Foundation
Abstract: The present invention provides methods, compositions, and kits for detecting the presence or absence of target sequences in a sample, where the sample also contains interfering sequences that are similar or identical to the target sequences. In particular, the present invention provides blocker oligonucleotides that at least partially inhibit the formation of invasive cleavage structures with the interfering sequences but do not substantially inhibit the formation of invasive cleavage structures with the target sequences.
Type:
Grant
Filed:
January 27, 2009
Date of Patent:
April 26, 2011
Assignee:
Third Wave Technologies, Inc.
Inventors:
Poonam Agarwal, Robert W. Kwiatkowski, Jr.
Abstract: The present invention relates to molecular constructs and methods of their use in detecting biochemical reactions. In particular, the invention relates to a molecular construct having a capture portion and a substrate portion, where the capture portion isolates the construct from a sample medium, and the substrate portion enables the construct to be acted upon and undergo a physical change which can be detected and measured. These molecular constructs may be used in diagnostic assays, genetic screening for potential risks of certain diseases in individuals, and drug discovery and toxicogenomics, using high throughput screening of compounds.
Abstract: Methods of detecting one or more nucleic acids from whole blood or plasma are provided. The nucleic acids are captured on a solid support and detected. Compositions, kits, and systems related to the methods are also described.