Abstract: Provided herein is reagent mixture comprising multiplexed amplification reagents and flap assay reagents for detecting, in a single reaction, mutant copies of the KRAS gene that contain any of the 34A, 34C, 34T, 35A, 35C, 35T or 38A point mutations. Methods that employ the reagent mix and kits for performing the same are also provided.
Type:
Grant
Filed:
March 10, 2020
Date of Patent:
June 7, 2022
Assignee:
Exact Sciences Development Company, LLC
Inventors:
Rebecca Oldham-Haltom, Hatim Allawi, Hongzhi Zou, Michael J. Domanico, Graham P. Lidgard
Abstract: Described herein are methods and compositions that provide highly efficient nucleic acid amplification. In some embodiments, this allows a 3-fold or greater increase of amplification product for each amplification cycle and therefore increased sensitivity and speed over conventional PCR. Modified bases can be employed in primers to provide this base-3 or greater amplification with satisfactory PCR cycle times, which are improved, as compared to those observed in the absence of modified bases.
Abstract: A microfluidic device includes a plurality of reaction wells; and a plurality of solid supports, and each of the solid supports has a reagent attached thereto. The reagent is attached to the solid support via a labile reagent/support bond such that the reagent is configured to be cleaved from the support via a cleaving operation.
Type:
Grant
Filed:
February 28, 2017
Date of Patent:
May 31, 2022
Assignee:
The University of North Carolina at Chapel Hill
Inventors:
John Michael Ramsey, William Henley, Emily Oblath
Abstract: The present disclosure provides methods for determining the ploidy status of a chromosome in a gestating fetus from genotypic data measured from a mixed sample of DNA comprising DNA from both the mother of the fetus and from the fetus, and optionally from genotypic data from the mother and father. The ploidy state is determined by using a joint distribution model to create a plurality of expected allele distributions for different possible fetal ploidy states given the parental genotypic data, and comparing the expected allelic distributions to the pattern of measured allelic distributions measured in the mixed sample, and choosing the ploidy state whose expected allelic distribution pattern most closely matches the observed allelic distribution pattern. The mixed sample of DNA may be preferentially enriched at a plurality of polymorphic loci in a way that minimizes the allelic bias, for example using massively multiplexed targeted PCR.
Type:
Grant
Filed:
November 4, 2014
Date of Patent:
May 3, 2022
Assignee:
Natera, Inc.
Inventors:
Matthew Rabinowitz, George Gemelos, Milena Banjevic, Allison Ryan, Zachary Demko, Matthew Hill, Bernhard Zimmermann, Johan Baner
Abstract: The present disclosure provides methods for determining the ploidy status of a chromosome in a gestating fetus from genotypic data measured from a sample of DNA from the mother of the fetus and from the fetus, and from genotypic data from the mother and optionally also from the father. The ploidy state is determined by using a joint distribution model to create a set of expected allele distributions for different possible fetal ploidy states given the parental genotypic data, and comparing the expected allelic distributions to the pattern of measured allelic distributions measured in the mixed sample, and choosing the ploidy state whose expected allelic distribution pattern most closely matches the observed allelic distribution pattern. In an embodiment, the mixed sample of DNA may be preferentially enriched at a plurality of polymorphic loci in a way that minimizes the allelic bias.
Type:
Grant
Filed:
February 12, 2014
Date of Patent:
April 19, 2022
Assignee:
Natera, Inc.
Inventors:
Matthew Rabinowitz, George Gemelos, Milena Banjevic, Allison Ryan, Zachary Demko, Matthew Hill, Bernhard Zimmerman, Johan Baner
Abstract: The present disclosure relates to methods, compositions, and kits for generating a library of tagged nucleic acid fragments without using PCR amplification, including methods and compositions for fragmenting and tagging nucleic acids (e.g., DNA) using transposome complexes immobilized on solid support.
Type:
Grant
Filed:
May 3, 2021
Date of Patent:
April 19, 2022
Assignee:
Illumina Cambridge Limited
Inventors:
Andrew Slatter, Esther Musgrave-Brown, Susan C. Verity, Niall Anthony Gormley
Abstract: The present invention provides a molecular marker C42257 for rapidly identifying genetic sex of Marsupenaeus japonicus and applications thereof. The nucleotide sequence of the molecular marker C42257 is shown in SEQ ID NO:1. The nucleotide sequences of the primer pair of molecular marker C42257 used for detection are C42257F: GGGGGACAA ACAGAGACA (SEQ ID NO: 2) and C42257R: ATCGGGGTGGATTTAGAA (SEQ ID NO: 3), respectively. The molecular marker C42257 is not affected by the tissue specificity of M. japonicus and the environment, and the steps are simple and the results are obvious. The genomic DNA of M. japonicus and the primer pair are subjected to PCR reaction and electrophoresis detection. If a target band appears at 183 bp, it is a female M. japonicus. The molecular marker can also promote the establishment of the seedling breeding technology of all-female or high-female-ratio M. japonicus. Therefore, it has a broad application prospect.
Type:
Grant
Filed:
September 2, 2021
Date of Patent:
April 19, 2022
Assignee:
Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences
Abstract: The present invention relates to the detection of a target nucleic acid sequence by a PTOCE (PTO Cleavage and Extension) assay. The present invention detects a target nucleic acid sequence in which the PTO (Probing and Tagging Oligonucleotide) hybridized with the target nucleic acid sequence is cleaved to release a fragment and the fragment is hybridized with the CTO (Capturing and Templating Oligonucleotide) to form an extended duplex, followed by detecting the presence of the extended duplex. The extended duplex provides signals (generation, increase, extinguishment or decrease of signals) from labels indicating the presence of the extended duplex and has adjustable Tm value, which are well adoptable for detection of the presence of the target nucleic acid sequence.
Abstract: Provided herein is technology relating to the amplification-based detection of bisulfite-treated DNAs and particularly, but not exclusively, to methods and compositions for multiplex amplification of low-level sample DNA prior to further characterization of the sample DNA. The technology further provides methods for isolating DNA from blood or blood product samples, e.g., plasma samples.
Abstract: A method for analyzing planar sample is provided. In some cases the method comprises: (a) incubating the planar sample with a capture agent that is linked to an oligonucleotide, wherein the capture agent specifically binds to complementary sites in the planar sample; (b) reading a fluorescent signal caused by extension of a primer that is hybridized to the oligonucleotide, using fluorescence microscopy. Several implementations of the method, and multiplexed versions of the same, are also provided.
Type:
Grant
Filed:
January 4, 2019
Date of Patent:
April 12, 2022
Assignee:
THE BOARD OF TRUSTEES OF THE LELAND STANFORD JUNIOR UNIVERSITY
Inventors:
Nikolay Samusik, Garry P. Nolan, Yury Goltsev
Abstract: A method of identifying a polynucleic acid (PNA) is presented, including the steps of providing a PNA; modifying one or more nucleobases of the PNA by addition or removal of a hydrogen bonding partner, thereby altering the base pairing capacity of the one or more nucleobases; base pairing a complementary nucleic acid to the PNA, including base pairing to at least one modified nucleobase; identifying the sequence of the complementary nucleic acid at least at the position that is complementary to at least one modified nucleobase.
Type:
Grant
Filed:
April 13, 2018
Date of Patent:
April 12, 2022
Assignee:
IMBA—INSIIIUT FÜR MOLEKULARE BIOTECHNOLOGIE GMBH
Inventors:
Stefan L. Ameres, Brian Reichholf, Veronika A. Herzog, Johannes Zuber, Matthias Muhar
Abstract: Provided herein, among other things, is an automatable procedure that employs in vitro directed evolution to create DNA sequences that encode a ligand-responsive ribozyme and which, when transcribed, can control expression of genes they are coupled to. The method also allows creation of functional RNA sequences that bind target molecules, without requiring any modification or immobilization of the target.
Type:
Grant
Filed:
May 27, 2020
Date of Patent:
April 5, 2022
Assignee:
THE BOARD OF TRUSTEES OF THE LELAND STANFORD JUNIOR UNIVERSITY
Abstract: Modified nucleic acid adapters are provided that collectively provide a mixture of nucleotides at the 3? end of 5? adapters and at the 5? end of 3? adapters such that at least one adapter in each set has any given nucleotide at position 1, i.e., the nucleotide position available for ligation to a small RNA, and has any given nucleotide at position 2 adjacent to position 1 for use in overcoming bias during nucleic acid manipulation, such as small RNA characterization and/or profiling by, e.g., deep sequencing, along with methods for use of the modified adapters in small RNA characterization. The modified adapters have at least two mixed nucleotides at the adapter terminus to be ligated to a nucleic acid such as a small RNA.
Type:
Grant
Filed:
February 7, 2020
Date of Patent:
April 5, 2022
Assignee:
Mount Sinai School of Medicine
Inventors:
Ravi Sachidanandam, Anitha Jayaprakash, Brian Brown
Abstract: In some aspects, the present disclosure provides methods for enriching amplicons, or amplification products, comprising a concatemer of at least two or more copies of a target polynucleotide. In some embodiments, a method comprises sequencing the amplicons comprising at least two or more copies of a target polynucleotide. In some embodiments, the target polynucleotides comprise sequences resulting from chromosome rearrangement, including but not limited to point mutations, single nucleotide polymorphisms, insertions, deletions, and translocations including fusion genes. In some aspects, the present disclosure provides compositions and reaction mixtures useful in the described methods.
Abstract: The invention relates to a method for in vitro investigating mitochondrial replication dysfunction in a biological sample removed from a subject susceptible of suffering from physiological ageing or physiopathological conditions related to physiological ageing, or physiopathological ageing or associated symptoms or conditions, in particular premature ageing or accelerated ageing, or of a progeroid syndrome, such as Cockayne syndrome (CS), or neurodegenerative disorders or symptoms thereof, in which the levels of at least one species selected in the group of: POLG1 protein, POLG1 RNA, POLG2 protein, protease(s) which have POLG as a target, in particular serine protease(s) such as HTRA3 protein, HTRA2 protein and, HTRA3 RNA or HTRA2 RNA, or any combination of these species, are investigated.
Type:
Grant
Filed:
June 20, 2019
Date of Patent:
March 29, 2022
Assignees:
INSTITUT PASTEUR, CENTRE NATIONAL DE LA RECHERCHE SCIENTIFIQUE, COMMISSARIAT A L'ENERGIE ATOMIOUE ET AUX ENERGIES ALTERNATIVES
Abstract: There is disclosed a composition of an aqueous solution comprising, consisting or consisting essentially of a flap endonuclease, a bulking agent and an organic buffer, wherein the aqueous solution has an inorganic salt concentration of 5 mM or less and wherein the composition is substantially free of glycerol.
Type:
Grant
Filed:
May 18, 2018
Date of Patent:
March 29, 2022
Assignee:
Gen-Probe Incorporated
Inventors:
Patrick Peterson, Tony Luu, Matthias Jost
Abstract: The present invention discloses a primer combination for detecting 9 NA subtype AIVs and an application thereof. The present invention protects a primer combination comprising 10 primer pairs (20 primers), and the 20 primers are set forth in SEQ ID NO: 1 to SEQ ID NO: 20 of the Sequence Listing in sequence. When used for identifying 9 NA subtypes, a GeXP detection method in combination with the primer combination provided by the present invention has the advantages of high throughput, fast, time-saving and labor-saving. The present invention provides an effective method for rapid identification of 9 different NA subtype AIVs and has a broad application prospect.
Type:
Grant
Filed:
April 24, 2019
Date of Patent:
March 22, 2022
Assignee:
GUANGXI VETERINARY RESEARCH INSTITUTE
Inventors:
Zhixun Xie, Sisi Luo, Jiaoling Huang, Zhiqin Xie, Liji Xie, Minxiu Zhang, Meng Li, Sheng Wang, Dan Li, Tingting Zeng, Yanfang Zhang, Qing Fan, Xianwen Deng
Abstract: High throughput personal genomic testing has created a need for robust quality control mechanisms to track sample identity, reagent integrity, and other factors with significant influence on assay performance. A method of massively parallel sequencing using an accompanying barcoded molecular standard enables one to track nucleic acid analytes to identify them by project, lot, batch, or patient. The molecular standard contains sequences present in the analyte, allowing it to be processed simultaneously without any other additional reagents. Within the molecular standard, a calibrator sequence permits assessment of fidelity of sequence determination. Additional sequences in the molecular standard may be used to manipulate the molecular standard separate from the analyte. The molecular standard can be used to benchmark sequencing platforms and assess error rates.
Type:
Grant
Filed:
February 14, 2019
Date of Patent:
March 8, 2022
Assignee:
THRIVE EARLIER DETECTION CORP.
Inventors:
Isaac A. Kinde, Howard B. Kaufman, Leonardo Hagmann