Abstract: A method is provided comprising the following steps: (a) treating a nucleic acid with bisulfite to convert non-methylated cytosines in the nucleic acid into uracils while leaving methylated cytosines unchanged to form a treated nucleic acid strand that is part of two joined nucleic acid strands; (b) ligating a first adapter to a 3? end of the treated nucleic acid strand, the first adapter having a first protruding random sequence that least 3 bases long and that acts as a splint for the two joined nucleic acid strands; (c) ligating a second adapter to a 5? end of the once adapter ligated nucleic acid strand, the second adapter having a second protruding random sequence at least 3 bases long and that acts as a splint for the two joined nucleic acid strands; and (d) performing PCR amplification on the twice ligated nucleic acid strand.
Abstract: Very simple, highly sensitive detection or quantification of target nucleic acids of interest has been achieved by: hybridizing mask oligonucleotides to regions in a single-stranded region of a nucleic acid to be assayed between which a region to be hybridized by an oligonucleotide probe is positioned, thereby opening the probe-hybridizing region and keeping the single-stranded region of the target nucleic acid stable, and then subjecting this nucleic acid having the single-stranded region to nucleic acid chromatography.
Abstract: The present invention provides a method for genomic profiling of DNA 5-methylcytosine and 5-hydroxymethylcytosine, comprising the following steps: (1) DNA purification and fragmentation pretreatment: the target DNA is extracted and then broken to an average of 50 nucleotides to 10,000 nucleotides in length; (2) the repair of trace amount of DNA and the ligation thereof to the adaptor: the pre-treated DNA fragments are repaired and ligated with the sequencing adaptor required for the second-generation sequencing, (3) covalently labeling 5-methylcytosine and 5-hydroxymethylcytosine, (4) solid-phase enrichment of the labeled DNA fragments having cytosine with 5-position modification; (5) the PCR amplification of the solid-phase enriched DNA fragments, the PCR product is obtained and purified to obtain a library for the second-generation sequencing, after mapping the sequencing reads to the genome, the distribution map of the cytosine with 5-position modification in the DNA sample could be generated.
Abstract: Methods for purifying RNA from a sample, comprising one or more steps of tangential flow filtration, hydroxyapatite chromatography, core bead flow-through chromatography, or any combinations thereof. These techniques are useful individually, but show very high efficiency when used in combination, or when performed in particular orders. The methods can purify RNA in a highly efficient manner without unduly compromising potency or stability, to provide compositions in which RNA is substantially cleared of contaminants. Moreover, they can be performed without the need for organic solvents.
Type:
Grant
Filed:
March 13, 2014
Date of Patent:
October 26, 2021
Assignee:
GLAXOSMITHKLINE BIOLOGICALS SA
Inventors:
Francesco Berlanda Scorza, Yingxia Wen, Andrew Geall, Frederick Porter
Abstract: The invention relates to methods for conducting solid-phase binding assays. One example is an assay method having improved analyte specificity where specificity is limited by the presence of non-specific binding interactions.
Type:
Grant
Filed:
December 5, 2018
Date of Patent:
October 19, 2021
Assignee:
Meso Scale Technologies, LLC
Inventors:
Eli N. Glezer, Sudeep Kumar, Pankaj Oberoi, George Sigal, Michael Tsionsky
Abstract: The present invention comprises methods and systems to profile individual chromosomes using target-specific DNA probes in biological samples. The invention relates to generation of chromosome profiles either singly or in combination (multiplex). The invention can refer to the generation of chromosome profiles using target-specific DNA probes for various biological samples such as cell free DNA from the peripheral blood of a pregnant woman or from a cancer patient. The invention further involving generation of chromosome profiles using target-specific DNA probes for individual intact cells from the peripheral blood of a pregnant woman, from a cancer patient or from an embryo created using artificial reproductive technologies. The invention further involving detection of target-specific DNA hybridizations through direct fluorescence by special spectral filters or fluorescence intensity by fluorimeters.
Abstract: Disclosed are methods for lysis of cells that combine three lysis steps—(1) heat, (2) detergent and (3) base—into a single step and that can be completed in a short period of time, e.g., a few minutes. The methods combine a normally incompatible detergent and base lysis, allow for simplified removal of detergent after lysis, and importantly, limits damage to DNA, such as shearing, that typically results from separate application of conventional lysis methods, yielding improved quality and quantities of genomic DNA (gDNA).
Abstract: The present disclosure encompasses methods of error corrected sequencing (ECS) that enable detection of very rare mutations well below the error rate of convention next generation sequencing (NGS). Further, the methods disclosed herein enable multiplex targeting of genomic DNA.
Abstract: The present invention features a method and kit for isolating microvesicles or extracting microvesicle nucleic acids from a biological sample by using a control particle. The present invention provides control particles that are viruses or virus-like particles, such as bacteriophages, that contain control nucleic acids that can be detected to assess the accuracy, reliability, and efficiency of the microvesicle isolation or nucleic acid extraction steps. The methods described herein may further comprise the analysis of the presence, absence, or level of at least one biomarker associated with a disease or medical condition for diagnosing, prognosing, or monitoring the disease or medical condition.
Type:
Grant
Filed:
August 30, 2013
Date of Patent:
October 5, 2021
Assignee:
Exosome Diagnostics, Inc.
Inventors:
Mikkel Noerholm, Susan Belzer, Charlotte Romain, Johan Karl Olov Skog, Leileata M. Russo, Wayne Comper
Abstract: Disclosed herein are methods of detecting a target RNA, methods of diagnosing an individual with a disease or condition when a target RNA associated with the disease or condition is detected, and methods of conveying via a communication medium data from the detection of a target RNA.
Abstract: A method for lysing cells is disclosed. The method includes stirring cells with a magnetic stir element in the presence of a plurality of cell lysis beads at a speed sufficient to lyse the cells. Also disclosed is a device for lysing cells. The device includes a container having a magnetic stir element and a plurality of cell lysis beads disposed therein. The container is dimensioned to allow rotation of the magnetic stir element inside the container.
Abstract: The invention generally relates to a molecular classification of disease predisposition and particularly to molecular markers for cancer predisposition and methods of use thereof.
Type:
Grant
Filed:
December 5, 2014
Date of Patent:
August 31, 2021
Assignee:
Myriad Genetics, Inc.
Inventors:
Kirsten Timms, Brian Allen, Anne-Renee Hartman
Abstract: The present disclosure provided methods and compositions for nucleic acid sequencing. In particular, the disclosure provides for detection of multiple different nucleotides in a sample utilizing fewer detection moieties than the number of nucleotides being detected and using two imaging events per sequencing cycle.
Abstract: The present invention generally relates to systems and methods for imaging or determining nucleic acids, for instance, within cells. In some embodiments, the transcriptome of a cell may be determined. Certain embodiments are directed to determining nucleic acids, such as mRNA, within cells at relatively high resolutions. In some embodiments, a plurality of nucleic acid probes may be applied to a sample, and their binding within the sample determined, e.g., using fluorescence, to determine locations of the nucleic acid probes within the sample. In some embodiments, codewords may be based on the binding of the plurality of nucleic acid probes, and in some cases, the codewords may define an error-correcting code to reduce or prevent misidentification of the nucleic acids. In certain cases, a relatively large number of different targets may be identified using a relatively small number of labels, e.g., by using various combinatorial approaches.
Type:
Grant
Filed:
July 29, 2015
Date of Patent:
August 24, 2021
Assignee:
President and Fellows of Harvard College
Inventors:
Xiaowei Zhuang, Kok-Hao Chen, Alistair Boettiger, Jeffrey R. Moffitt, Siyuan Wang
Abstract: This disclosure provides methods and compositions for detecting intramolecular and intermolecular interactions, such as protein-protein interactions. These methods detect such interactions at sub-diffraction distances, and thus are referred to as super-resolution detection and imaging methods.
Type:
Grant
Filed:
August 5, 2016
Date of Patent:
August 17, 2021
Assignee:
President and Fellows of Harvard College
Inventors:
Juanita M. Lara Gutierrez, Maier S. Avendano Amado, Peng Yin
Abstract: The present invention is directed to a method for immobilizing a nucleic acid molecule on a solid support and to a use of a combination of a first nucleic acid immobilized primer linked to a solid support and a second immobilized primer linked to said solid support in said method.
Abstract: The present invention relates to the detection of a target nucleic acid sequence by a PCEC (PTO Cleavage and Extension-Dependent Cleavage) assay. The present invention is characterized by generating a cleavage site for a nucleolytic enzyme on the extended duplex of which the formation is dependent on the presence of a target nucleic acid sequence. The present invention detects the occurrence of the cleavage of the extended duplex, thereby determining the presence of the target nucleic acid sequence.
Abstract: In a first aspect, the invention concerns a method for detecting or quantifying DNA methylation at a locus. In one embodiment, a methylation-sensitive endonuclease is formulated together with a polymerase enzyme in an appropriate reaction mixture such that amplification of DNA occurs in a methylation specific manor. Quantitative DNA amplification at selected loci can be used to determine the level of methylation. Kits and reagents for performing such methods are also provided.
Abstract: The present invention relates to methods of imaging template hybridisation for estimating cluster numbers prior to solid phase amplification and sequencing. More particularly, an initial round of imaging is carried out at the single molecule template hybridisation stage which allows a general estimation of cluster numbers prior to clusters being formed. Amplification of the signal allows single molecule imaging to be carried out using standard sequencing imaging apparatus.
Type:
Grant
Filed:
June 11, 2015
Date of Patent:
July 13, 2021
Assignee:
Illumina Cambridge Limited
Inventors:
Isabelle Marie Julia Rasolonjatovo, Jonathan Mark Boutell, Vincent Peter Smith, Roberto Rigatti