Abstract: The present disclosure relates to materials and methods for spatially analyzing nucleic acids that have been fragmented with a transposase enzyme, alone or in combination with other types of analytes
Type:
Application
Filed:
August 27, 2019
Publication date:
October 12, 2023
Applicant:
10x Genomics, Inc.
Inventors:
Michael Schnall-Levin, Michael Ybarra Lucero, Tarjei Sigurd Mikkelsen, Patrik Stahl, Jonas Frisen, Maja Marklund, Enric Llorens Bobadilla, Eswar Prasad Ramachandran Iyer, Lucas Frenz, Augusto Manuel Tentori, Rajiv Bharadwaj
Abstract: Provided herein in some aspects are methods, compositions, kits, and systems for performing multiplexed single-cell analysis on an in situ platform, providing alternatives which have a higher cell throughput and/or lower cost per cell compared to current single-cell analysis techniques. In some embodiments, the methods disclosed herein comprise using labeling agents that comprise sample-specific barcodes and/or cell feature specific barcodes (e.g., analyte specific barcodes) to label single-cell populations, immobilizing the labeled cells, and performing in situ detection of the labeling agents and/or other features including cellular analytes of the labeled cells.
Abstract: Disclosed herein are systems for receiving a biological sample for analysis and methods for preparing and/or analyzing a biological sample. In various embodiments, a system includes a first layer comprising a first region configured to receive a sample, a gasket disposed on the first layer such that the gasket surrounds the first region, and a housing layer. The first layer is disposed between the gasket and the housing layer. The system further includes a clamp configured to apply a force on the gasket to form a liquid tight seal against the first layer.
Abstract: The present disclosure relates in some aspects to methods for analyzing a target nucleic acid in a biological sample. In some aspects, provided herein are methods and compositions for improving the specificity of ligation in situ in biological samples, as well as in single cell analysis and spatial applications of RNA templated ligation reactions.
Abstract: Provided herein are methods of determining a surgical margin and the site and size of a tissue to be resected from a subject, and methods of use thereof.
Abstract: The present application provides methods for detecting a nucleic acid molecule involving the use of a signal code sequence which corresponds to said nucleic acid molecule and a plurality of labelled detection probes which yield signals which make up the signal code sequence. In particular, the invention provides a sequential barcoding and decoding scheme which utilises a sequencing-by-hybridisation (SBH) strategy to sequence and decode a nucleotide barcode sequence, and to differentiate the nucleotide barcode sequence from other nucleotide barcode sequences. In an extension of the method, the application also provides a new coding scheme for providing a target nucleic acid with a detectable “colour” (or similar signal)-based code.
Abstract: Provided herein are methods of determining a location of an analyte in a biological sample and devices that include a plurality of wells, where a well of the plurality of wells comprises a surface comprising a plurality of capture probes.
Type:
Application
Filed:
September 16, 2021
Publication date:
October 5, 2023
Applicant:
10x Genomics, Inc.
Inventors:
Stefania Giacomello, Paulius Mielinis, Marlon Stoeckius
Abstract: Methods and systems for sample preparation techniques that allow amplification (e.g., whole genome amplification) and sequencing of chromatin accessible regions of single cells are provided. The methods and systems generally operate by forming or providing partitions (e.g., droplets) including a single biological particle and a single bead comprising a barcoded oligonucleotide. The preparation of barcoded next-generation sequencing libraries prepared from a single cell is facilitated by the transposon-mediated transposition and fragmentation of a target nucleic acid sequence. The methods and systems may be configured to allow the implementation of single-operation or multi-operation chemical and/or biochemical processing within the partitions.
Abstract: The present disclosure provides electrophoretic systems, methods and compositions for spatial analysis, which can serve to magnify or demagnify spatial resolution of analytes of interest that are captured using electrophoresis. Some implementations can use a diverging or converging electric field in an electrophoretic capture system. Such a divergent or convergent electric field, as opposed to a parallel electric field, can be generated by, for example, utilizing different sizes of electrodes associated with or imbedded in substrates. Also provided herein are electrophoretic systems, methods and compositions for spatial analysis, which can serve to selectively migrate one or more analytes from a region of interest in the biological sample for capture using electrophoresis.
Type:
Grant
Filed:
March 4, 2021
Date of Patent:
September 26, 2023
Assignee:
10x Genomics, Inc.
Inventors:
Hanyoup Kim, Augusto Manuel Tentori, Rajiv Bharadwaj
Abstract: Provided herein are methods of identifying a location of an RNA in a sample that include: (a) contacting the sample with an array comprising capture probes, where a capture probe comprises a capture domain and a spatial barcode; (b) releasing the RNA from the sample; (c) extending a 3? end of the capture probe using the capture domain-bound RNA as a template; (d) generating nick(s) in the extended capture probe-hybridized RNA and performing random-primed DNA synthesis; (e) performing end repair on the second strand DNA molecule; (f) adding a single adenosine nucleotide to the 3? end of the extended capture probe; (g) ligating a double-stranded sequencing adaptor to the double-stranded DNA product; and (h) determining all or a part of the sequence of the RNA, and the sequence of the spatial barcode, or complements thereof, and using the determined sequences to identify the location of the RNA in the sample.
Abstract: The present disclosure relates in some aspects to methods, probes, and kits for detection of a target analyte in a sample. In some embodiments, disclosed herein are methods in which a biological sample is contacted with a pre-formed detectable probe comprising a concatemeric region, e.g., a rolling circle amplification (RCA) product, comprising multiple copies of a unit sequence. Also disclosed herein are related probes and kits.
Abstract: Provided are methods for determining a location of a target nucleic acid in a biological sample including: disposing the biological sample onto an array including a plurality of capture probes, where a first capture probe includes a first spatial barcode and a capture domain and a second capture probe includes a second spatial barcode and the capture domain. The second capture probe is not covered by the biological sample on the array and is contacted with a solution comprising TdT and one or more dideoxynucleotides, such that a dideoxynucleotide is incorporated into the second capture domain. Target nucleic acids are captured by the first capture probe, and the sequence of the first spatial barcode or a complement thereof and all or a portion of a sequence of the target nucleic acid, or a complement thereof, are used to determine the location of the target nucleic acid in the biological sample.
Abstract: The present disclosure relates to compositions and methods for generating capture probes on a substrate for identifying the location of analytes in a biological sample.
Abstract: A discrete attribute value dataset is obtained that is associated with a plurality of probe spots each assigned a different probe spot barcode. The dataset comprises spatial projections, each comprising images of a biological sample. Each image includes a corresponding plurality of discrete attribute values for the probe spots. Each such value is associated with a probe spot in the plurality of probes spots based on the probe spot barcodes. The dataset is clustered using the discrete attribute values, or dimension reduction components thereof, for a plurality of loci at each respective probe spot across the images of the projections thereby assigning each probe spot to a cluster in a plurality of clusters. Morphological patterns are identified from the spatial arrangement of the probe spots in the various clusters.
Type:
Grant
Filed:
October 18, 2022
Date of Patent:
September 12, 2023
Assignee:
10X GENOMICS, INC.
Inventors:
Jeffrey Clark Mellen, Jasper Staab, Kevin J. Wu, Neil Ira Weisenfeld, Florian Baumgartner, Brynn Claypoole
Abstract: In some aspects disclosed herein are methods and compositions for detecting analytes such as a plurality of nucleic acid molecules in a biological sample, said method comprising generating and analyzing a first detectable signal and a second detectable signal corresponding to a first target sequence and a second target sequence, respectively. The first and second target sequences can be present and/or expressed at different levels in the biological sample.
Abstract: The present disclosure relates in some aspects to methods and compositions for analysis of a target nucleic acid, such as in situ detection of a region of interest in a polynucleotide in a tissue sample. In some aspects, provided herein are methods and compositions for detecting a region of interest in a target nucleic acid. In some aspects, provided herein are circularizable probes and oligonucleotide probes for analyzing a target nucleic acid, in which corresponding hybridization regions between the circularizable probe and the oligonucleotide probe comprise a barcode region comprising one or more barcodes. In some aspects, the oligonucleotide probe and/or the circularizable probe comprise one or more modifications.
Abstract: The present invention is directed to methods, compositions and systems for capturing and analyzing sequence information contained in targeted regions of a genome. Such targeted regions may include exomes, partial exomes, introns, combinations of exonic and intronic regions, genes, panels of genes, and any other subsets of a whole genome that may be of interest.
Type:
Grant
Filed:
October 6, 2020
Date of Patent:
August 29, 2023
Assignee:
10X GENOMICS, INC.
Inventors:
Mirna Jarosz, Michael Schnall-Levin, Serge Saxonov, Benjamin J. Hindson, Xinying Zheng
Abstract: The present disclosure provides compositions, methods, systems, and devices for polynucleotide processing and analyte characterization from a single cell. Such polynucleotide processing may be useful for a variety of applications. The compositions, methods, systems, and devices disclosed herein generally describe barcoded oligonucleotides, which can be bound to a bead, such as a gel bead, useful for characterizing one or more analytes including, for example, protein (e.g., cell surface or intracellular proteins) and chromatin (e.g., accessible chromatin).
Type:
Grant
Filed:
June 9, 2020
Date of Patent:
August 29, 2023
Assignee:
10X GENOMICS, INC.
Inventors:
Jason Bell, Geoffrey McDermott, Francesca Meschi, Katherine Pfeiffer, Michael Schnall-Levin, Xinying Zheng
Abstract: Provided herein are methods, compositions, and kits for preparing biological samples for multiplex spatial gene expression and proteomic analysis, such as determining a location of a nucleic acid analyte and a protein analyte in a biological sample.
Type:
Grant
Filed:
July 18, 2022
Date of Patent:
August 29, 2023
Assignee:
10x Genomics, Inc.
Inventors:
Jennifer Chew, Joseph Francis Shuga, Patrick Roelli, Denise Cheung, Alexander Hermes, Yifeng Yin, Naishitha Anaparthy, Ryo Hatori, Marlon Stoeckius, Rapolas Spalinskas, Anna-Maria Katsori, Cedric Uytingco, Mesruh Turkekul, David Sukovich, Christina Galonska, Aleksandra Jurek, Octavian Marian Bloju