Abstract: The present disclosure provides a solid phase method of making oligonucleotides via sequential coupling cycles including at least one coupling of a dinucleotide dimer subunit to a free 3?-terminal group of a growing chain. The oligonucleotides include at least two nucleoside subunits joined by a N3??P5? phosphoramidate linkage. The method may include the steps of (a) deprotecting the protected 3? amino group of a terminal nucleoside attached to a solid phase support, said deprotecting forming a free 3? amino group; (b) contacting the free 3? amino group with a 3?-protected amino-dinucleotide-5?-phosphoramidite dimer in the presence of a nucleophilic catalyst to form an internucleoside N3??P5? phosphoramidite linkage; and (c) oxidizing (e.g., sulfurizing) the linkage. The compositions produced by the subject methods may include a reduced amount of one or more (N?x) oligonucleotide products. Also provided are pharmaceutical compositions including the subject oligonucleotide compositions.
Abstract: In an example, a flow cell includes a substrate, a selectively removable porous molecular network on the substrate and defining exposed substrate regions, and sequencing surface chemistry on at least some of the exposed regions. The sequencing surface chemistry is selected from the group consisting of i) an activated pad, a polymer layer attached to the activated pad, and a primer attached to the polymer layer; or ii) a nanostructure and an enzyme attached to the nanostructure.
Abstract: Provided herein are compositions, methods and systems relating to libraries of polynucleotides such that the libraries allow for accurate and efficient hybridization after binding to target sequences. Further provided herein are probes, blockers, additives, buffers, and methods that result in improved hybridization. Such compositions and methods are useful for improvement of Next Generation Sequencing applications, such as reducing off-target binding or reducing workflow times.
Type:
Grant
Filed:
October 1, 2019
Date of Patent:
August 22, 2023
Assignee:
Twist Bioscience Corporation
Inventors:
Ramsey Ibrahim Zeitoun, Siyuan Chen, Richard Gantt, Kristin D. Butcher, E. Hutson Chilton
Abstract: The present application describes compositions and methods for identifying and quantitating molecular targets within a cellular environment. Specifically, provided herein are compositions and methods for separately identifying and quantifying each of one or more molecular targets from a single cell. More specifically, provided herein are compositions and methods for separately identifying and quantifying the same molecular target from a single cell.
Abstract: Cleavable nucleotide analogs are provided. The nucleotide analog includes a nucleotide molecule attached to a cleavable moiety wherein the cleavable moiety comprises a protective group and/or a linker attached to a fluorophore. The cleavable moiety is linked to the oxygen atom of the 3?-OH of the pentose of the nucleotide molecule. The nucleotide analogs can be used in making polynucleotide molecules using template independent polymerases. The nucleotide analogs can act as reversible terminators during DNA sequencing by synthesis. The cleavage of the cleavable moiety restores a free 3?-OH functional group allowing growth of the polynucleotide molecule. The general structures as well as proposed synthetic schemes for the nucleotide analogs are also provided.
Type:
Grant
Filed:
November 30, 2017
Date of Patent:
August 15, 2023
Assignee:
President and Fellows of Harvard College
Inventors:
Kettner John Frederick Griswold, Jr., Richard E. Kohman, George M. Church, Jonathan Rittichier
Abstract: Compositions, methods, and systems are provided for fluorescent polymerase enzyme substrates comprising protein shields for improving enzyme photostability in single molecule real time sequencing. Fluorescent polymerase enzyme substrates of the invention have a protein shield between the fluorescent dye moieties and nucleotide moieties of the polymerase enzyme substrate. The polymerase enzyme substrates have a nucleotide component and a dye component, each attached to a protein. The attachments can be covalent. The protein can, for example, prevent the direct interaction of the fluorescent dye moiety with the enzyme when carrying out nucleotide synthesis, preventing photodamage to the enzyme. The polymerase enzyme substrates of the invention can have multiple dyes and multiple nucleotide moieties.
Type:
Grant
Filed:
April 14, 2021
Date of Patent:
August 8, 2023
Assignee:
Pacific Biosciences of California, Inc.
Inventors:
Keith Bjornson, Jeremiah Hanes, Erik Miller, Satwik Kamtekar, Lubomir Sebo, Louis Brogley
Abstract: Light harvesting luminescent multichromophores that are configured upon excitation to transfer energy to, and amplify the emission from, an acceptor signaling chromophore in energy-receiving proximity therewith are provided. Also provided are compositions for labelling a target. The labelling composition may include a donor light harvesting multichromophore and an acceptor signaling chromophore in energy-receiving proximity to the donor light harvesting multichromophore. Also provided is an aqueous composition for labelling a target, including: a donor light harvesting multichromophore; an acceptor signaling chromophore in energy-receiving proximity therewith; and a sensor biomolecule. Methods for using the subject compositions are also provided.
Type:
Grant
Filed:
July 31, 2019
Date of Patent:
August 8, 2023
Assignee:
THE REGENTS OF THE UNIVERSITY OF CALIFORNIA
Inventors:
Guillermo C. Bazan, Brent S. Gaylord, Shu Wang, Bin Liu
Abstract: Described herein are novel divalent nucleobases that each bind two nucleic acid strands, matched or mismatched when incorporated into a nucleic acid or nucleic acid analog backbone (a genetic recognition reagent, or genetic recognition reagent). In one embodiment, the genetic recognition reagent is a peptide nucleic acid (PNA) or gamma PNA (?PNA) oligomer. Uses of the divalent nucleobases and monomers and genetic recognition reagents containing the divalent nucleobases also are provided.
Type:
Grant
Filed:
December 28, 2021
Date of Patent:
August 1, 2023
Assignee:
Carnegie Mellon University
Inventors:
Danith H. Ly, Suresh Kumar Gopalsamy, Arunava Manna
Abstract: Provided are synthetic strands for nucleic acid sequencing. In some embodiments, the strands include a plurality of rotatable solid supports. The plurality of rotatable solid supports comprises solid supports each comprising on its surface a first moiety that binds to adenine (A), a second moiety that binds to cytosine (C), a third moiety that binds to guanine (G), a fourth moiety that binds to thymine (T), uracil (U), or both (T/U). Each of such solid supports further comprises on its surface a position marker that indicates the rotational position of the solid support, where the first, second, third, and fourth moieties are spaced about the circumference of the solid support. The solid supports enable hybridization of the synthetic strand to a nucleic acid. Also provided are methods of using the synthetic strands, as well as related compositions, kits, and nucleic acid sequencing systems.
Abstract: The present disclosure relates to methods, compositions, and kits for treating target nucleic acids, including methods and compositions for fragmenting and tagging nucleic acid (e.g., DNA) using transposome complexes bound to a solid support.
Type:
Grant
Filed:
January 4, 2021
Date of Patent:
July 25, 2023
Assignees:
Illumina, Inc.
Inventors:
Grace DeSantis, Stephen M. Gross, Jian-Sen Li, Natalie Morrell, Andrew Slatter, Kevin Shen, Samantha Snow
Abstract: A gene sequencing substrate and a method for manufacturing the same, and a gene sequencing device are provided. It belongs to the technical field of gene sequencing, and can solve the problem of high cost of the high-throughput sequencing chip in the prior art. The gene sequencing substrate of the present disclosure comprises a plastic material with concave structures as base substrate, and the concave structures serve as reaction cells. Since the base substrate has plasticity, the concave structures can be formed by a simple process to reduce the cost of the gene sequencing substrate. Meanwhile, a first protective layer may be provided on the inner wall of the concave structures for preventing the inner wall of the concave structures from being corroded by the reaction liquid.
Abstract: Methods and devices are provided herein for surfaces for de novo nucleic acid synthesis which provide for low error rates. In addition, methods and devices are provided herein for increased nucleic acid mass yield resulting from de novo nucleic acid synthesis.
Type:
Grant
Filed:
April 21, 2020
Date of Patent:
July 11, 2023
Inventors:
Pierre F. Indermuhle, Eugene P. Marsh, Andres Fernandez, William Banyai, Bill J. Peck
Abstract: The present invention concerns an oligonucleotide-functionalized hydrophobic polymer nanoparticle and method of its preparation. Said nanoparticle is a dye-loaded polymeric nanoparticle, and being functionalized by: (a) target-specific oligonucleotides, and/or (b) non-specific oligonucleotides.
Abstract: This disclosure relates to novel detergents for use in various procedures including, for example, nucleic acid amplification reactions such as polymerase chain reaction (PCR). Methods for preparing the modified detergents are also described.
Type:
Grant
Filed:
May 8, 2020
Date of Patent:
July 11, 2023
Assignee:
Life Technologies Corporation
Inventors:
Parul Angrish, Zhiwei Yang, Jonathan Wang
Abstract: The invention relates to a method for the specific detection of a microorganism or a group of microorganisms via in situ hybridisation by means of flow cytometry.
Type:
Grant
Filed:
August 16, 2018
Date of Patent:
June 20, 2023
Assignee:
Vermicon AG
Inventors:
Jiri Snaidr, Claudia Beimfohr, Peter Muhlhahn
Abstract: Provided are methods of producing product nucleic acids involving the use of oligonucleotides that are modified by the application of a stimulus. Aspects of such methods may include producing product nucleic acids using de-activatable oligonucleotides that are deactivated by a de-activating stimulus, as well as methods that may include producing product nucleic acids using activatable oligonucleotides that are activated by an activating stimulus and de-activatable oligonucleotides that are deactivated by a de-activating stimulus. Also provided are kits, compositions and devices that include de-activatable oligonucleotides or activatable and de-activatable oligonucleotides, e.g., for use in performing the methods as described herein.
Abstract: The novel intercalating fluorescent compounds of exemplary embodiments of the present invention for analyzing nucleic acids, etc. have excellent intercalating efficiency with nucleic acids such as DNA and RNA of biomaterials, and may not only continuously maintain fluorescence properties and efficiency, but also have excellent effects even in terms of storage stability such as temperature and moisture, etc. and biosafety. In addition, the fluorescent compounds have various advantages capable of being dissolved in distilled water, which is a solvent harmless to the human body, and being applied to a wide range of analysis without being limited to the analysis of specific cells and living tissues.
Type:
Grant
Filed:
December 19, 2020
Date of Patent:
June 6, 2023
Inventors:
Jin Woo Park, Ho Young Si, Eunae Jeong, Hyung Jun Yoon, Su Jung Jang
Abstract: Disclosed are PCR processes that transliterate target duplex DNA molecules containing 8 different nucleotide building blocks strategically arranged to allow efficient conversion of information in that target into DNA built from just four different nucleotides.
Abstract: Processes described here exploit the utility of mismatching nucleobase analogs to create long segments of natural DNA to be constructed after fragments containing unnatural nucleotides are transliterated in at least two cycles of polymerase-catalyzed copying, or alternatively complete PCR, by guided mismatching.