The present disclosure in some aspects relates to methods and compositions for confining molecules generated in a biological sample. In particular examples, rolling circle amplification (RCA) products are generated in a fixed biological sample which has been crosslinked prior to RCA, such that the three-dimensional crosslinked matrix generated prior to RCA confines the RCA products, and the RCA products are smaller than they would otherwise be in a fixed biological sample which has not been crosslinked prior to RCA. Confining the RCA products by generating them in a pre-crosslinked matrix in the biological sample may result in compaction of the RCA products and facilitate subsequent in situ analysis. The sample crosslinking can be performed after probe hybridization to nucleic acid molecules in the sample and before RCA.
The present disclosure relates generally to arrays for transcriptomics assays and methods of preparing the same. The disclosure provides arrays having features on a substrate and filling material in the spaces on the substrate that are not occupied by the features. Particularly, the disclosure provides arrays in which the features have a first refractive index at a wavelength and the filling material have a second refractive index at the same wavelength, such that the difference between the first refractive index and the second refractive index is no greater than about 10%. The disclosure also provides methods of preparing such arrays as well as methods of detecting biological analytes using said arrays.
G01N 21/25 - Couleur; Propriétés spectrales, c. à d. comparaison de l'effet du matériau sur la lumière pour plusieurs longueurs d'ondes ou plusieurs bandes de longueurs d'ondes différentes
Provided herein are methods, compositions, and kits for the spatial analysis of target nucleic acids in biological samples by their 5' end. An exemplary method includes: contacting a biological sample with a primer that hybridizes the target nucleic acid; hybridizing the primer to the target nucleic acid and extending using the target nucleic acid as a template to generate an extension product; incorporating a non-templated polynucleotide sequence including at least three nucleotides to the 3' end of the extension product; hybridizing the polynucleotide sequence of the extension product to a capture domain on an array including a plurality of capture probes that include a spatial barcode and the capture domain; and determining the spatial barcode sequence, or complement thereof, and all or a portion of the extension product, or complement thereof, and using the determined sequences to determine the location of the target nucleic acid in the biological sample.
The present disclosure provides compositions and methods for using fixed biological samples in partition-based assays. In at least one embodiment, the disclosure provides a composition comprising a fixed biological sample and an un-fixing agent contained in a partition, such as a discrete droplet. In some embodiments, the disclosure provides un-fixing agent compounds capable of catalytically cleaving crosslinks in fixed biological samples, particularly crosslinked nucleic acids, such as RNA.
G01N 33/53 - Tests immunologiques; Tests faisant intervenir la formation de liaisons biospécifiques; Matériaux à cet effet
C07D 213/04 - Composés hétérocycliques contenant des cycles à six chaînons, non condensés avec d'autres cycles, ne comportant qu'un atome d'azote comme unique hétéro-atome du cycle et avec au moins trois doubles liaisons entre chaînons cycliques ou entre chaînons comportant trois liaisons doubles ne comportant pas de liaison entre l'atome d'azote du cycle et un chaînon non cyclique ou ne comportant que des atomes d'hydrogène ou de carbone liés directement à l'atome d'azote du cycle
C07D 239/24 - Composés hétérocycliques contenant des cycles diazine-1, 3 ou diazine-1, 3 hydrogéné non condensés avec d'autres cycles comportant au moins trois liaisons doubles entre chaînons cycliques ou entre chaînons cycliques et chaînons non cycliques
C12N 9/48 - Hydrolases (3.) agissant sur les liaisons peptidiques, p.ex. thromboplastine, aminopeptidase de la leucine (3.4)
G01N 33/50 - Analyse chimique de matériau biologique, p.ex. de sang ou d'urine; Test par des méthodes faisant intervenir la formation de liaisons biospécifiques par ligands; Test immunologique
Provided herein are systems and methods for drug screening single cells from a sample. A method for drug screening may comprise treating various single cell samples with different drugs or drug combinations, labeling of treated cells with a labeling agent comprising a sample barcode, and processing the treated, labeled cells to generate nucleic acid libraries for sequencing. One or more processes of the methods described herein may be performed within a partition, such as a droplet or well.
C12Q 1/6876 - Produits d’acides nucléiques utilisés dans l’analyse d’acides nucléiques, p.ex. amorces ou sondes
C12Q 1/25 - Procédés de mesure ou de test faisant intervenir des enzymes, des acides nucléiques ou des micro-organismes; Compositions à cet effet; Procédés pour préparer ces compositions faisant intervenir des enzymes qui ne peuvent pas être classées dans les groupes
C12Q 1/6811 - Méthodes de sélection pour la production ou l’élaboration d’oligonucléotides spécifiques cibles ou de molécules de liaison
C12Q 1/6816 - Tests d’hybridation caractérisés par les moyens de détection
The present disclosure relates in some aspects to methods and compositions for quality control (e.g., assessing the level of fixation of a biological sample) and/or optimization of analyte detection in fixed samples. In some embodiments, the method can be used to process a fixed biological sample, the method comprising contacting the fixed biological sample with a nucleic acid stain and/or an actin stain; detecting an optical signal associated with the nucleic acid stain and/or an optical signal associated with the actin stain; comparing the detected optical signal(s) to one or more references to determine the quality of the biological sample. Single cell analysis, spatial array based analysis or in situ analysis can be performed using the processed biological sample.
G01N 33/50 - Analyse chimique de matériau biologique, p.ex. de sang ou d'urine; Test par des méthodes faisant intervenir la formation de liaisons biospécifiques par ligands; Test immunologique
G01N 33/58 - Analyse chimique de matériau biologique, p.ex. de sang ou d'urine; Test par des méthodes faisant intervenir la formation de liaisons biospécifiques par ligands; Test immunologique faisant intervenir des substances marquées
Provided in some aspects are methods for light-controlled in situ surface patterning of a substrate. Compositions such as nucleic acid arrays produced by the methods are also disclosed. Further provided in some aspects are methods for light-controlled in situ refinement of nucleic acid array feature boundaries. In some embodiments, a method disclosed herein comprises using photoresist for photocontrollable removal, blocking, and/or inactivation of nucleic acid molecules at feature boundaries. A large diversity of barcodes can be created in molecules on the substrate via sequential rounds of light exposure, hybridization, ligation, and further interceded and/or followed by one or more rounds of photocontrollable refinement of array feature boundaries, to improve spatial array resolution and sensitivity.
The present disclosure relates in some aspects to methods and compositions for in situ analysis involving catalytic de-crosslinking of biological samples.
The present disclosure relates generally to compositions, methods and systems for characterizing a biological particle, cell or cell nucleus, modified to include an exogenous insert, e.g., exogenous nucleic acids. The characterization may identify the insertion site of the exogenous nucleic acids in the biological particle's endogenous, e.g., genomic, nucleic acid sequences. Such characterization may be useful in the development of safer, more effective, modified cell biotherapeutics, e.g., cells modified to express chimeric antigen receptors.
C12Q 1/6818 - Tests d’hybridation caractérisés par les moyens de détection impliquant l’interaction de plusieurs marqueurs, p.ex. transfert d’énergie de résonance
C12N 5/00 - Cellules non différenciées humaines, animales ou végétales, p.ex. lignées cellulaires; Tissus; Leur culture ou conservation; Milieux de culture à cet effet
C12Q 1/6804 - Analyse d’acides nucléiques utilisant des immunogènes
C12Q 1/6837 - Couplage enzymatique ou biochimique d’acides nucléiques à une phase solide utilisant des réseaux de sondes ou des puces à sondes
C12Q 1/6876 - Produits d’acides nucléiques utilisés dans l’analyse d’acides nucléiques, p.ex. amorces ou sondes
G01N 33/50 - Analyse chimique de matériau biologique, p.ex. de sang ou d'urine; Test par des méthodes faisant intervenir la formation de liaisons biospécifiques par ligands; Test immunologique
G01N 33/543 - Tests immunologiques; Tests faisant intervenir la formation de liaisons biospécifiques; Matériaux à cet effet avec un support insoluble pour l'immobilisation de composés immunochimiques
14.
Methods and systems including flow and magnetic modules
Systems, and their methods of use, for sorting or separating magnetic particles are provided. A system having a magnetic module with features that mate with voids in a flow module exerts a magnetic field on magnetic particles to separate particles.
The present disclosure relates in some aspects to recombinant polymerases, variant PumA1 polymerases, and compositions thereof. Also provided herein are methods of using the recombinant polymerases and/or variant PumA1 polymerases for nucleic acid amplification (e.g., rolling circle amplification). In some aspects, the compositions and methods disclosed herein provide more robust amplification (e.g., RCA) reactions for improved in vitro and in situ analysis.
C12N 9/12 - Transférases (2.) transférant des groupes contenant du phosphore, p.ex. kinases (2.7)
C12N 15/63 - Introduction de matériel génétique étranger utilisant des vecteurs; Vecteurs; Utilisation d'hôtes pour ceux-ci; Régulation de l'expression
The present disclosure generally relates to methods and systems for engineering antibodies, and antigen-binding fragments thereof, to have altered characteristics. The present disclosure also provides a high-throughput method useful for identifying the engineered antibodies, or antigen-binding fragments thereof, that acquired the altered characteristics having performed the methods. These methods and systems have implications, for example, in the rapid development of biotherapeutics having desired and/or improved, e.g., affinity, specificity and/or activity-related, characteristics.
The present disclosure relates to materials and methods for spatially analyzing nucleic acids fragmented with a transposase enzyme in a biological sample.
A method for identifying framework regions and complementarity-determining regions in an amino acid sequence. The amino acid sequence is received. A plurality of candidate start positions is identified within the amino acid sequence for a start position for a selected region of interest. A score is generated for each candidate start position of the plurality of candidate start positions via analysis of a motif window that begins at each candidate start position. The start position for the selected region of interest is identified based on a candidate start position of the plurality of candidate start positions having a highest score.
G16B 20/30 - Détection de sites de liaison ou de motifs
G16B 30/00 - TIC spécialement adaptées à l’analyse de séquences impliquant des nucléotides ou des aminoacides
G16B 40/00 - TIC spécialement adaptées aux biostatistiques; TIC spécialement adaptées à l’apprentissage automatique ou à l’exploration de données liées à la bio-informatique, p.ex. extraction de connaissances ou détection de motifs
A sample holder includes a first member featuring a first retaining mechanism configured to retain a first substrate that includes a sample, a second member featuring a second retaining mechanism configured to retain a second substrate that includes a reagent medium, and an alignment mechanism connected to at least one of the first and second members, and configured to align the first and second members such that the sample contacts at least a portion of the reagent medium when the first and second members are aligned.
G02B 21/26 - Platines; Moyens de réglage pour celles-ci
G01N 33/483 - Analyse physique de matériau biologique
G01N 35/00 - Analyse automatique non limitée à des procédés ou à des matériaux spécifiés dans un seul des groupes ; Manipulation de matériaux à cet effet
G02B 21/34 - Lames de microscope, p.ex. montage d'échantillons sur des lames de microscope
G02B 21/36 - Microscopes aménagés pour la photographie ou la projection
The present disclosure relates in some aspects to recombinant Bacillus phage AP50 polymerases, modified AP50 polymerases, and compositions thereof. Also provided are methods of using the recombinant and/or modified AP50 polymerases for nucleic acid amplification (e.g., rolling circle amplification). In some aspects, the compositions and methods disclosed herein provide more robust amplification (e.g., RCA) reactions for improved in vitro and in situ analysis.
Methods and compositions for performing base-by-base sequencing in situ in a cell or tissue sample that minimize optical crowding are described. In some embodiments, a sequencing primer hybridizes to a priming site 3′ to an identifier sequence (e.g., a barcode sequence) in the sample such that the sequencing primer can be extended by a polymerase in a base-by-base fashion using the identifier sequence as a template. The sample can be contacted with nucleotides in a cyclic series of nucleotide incorporation or binding steps, and signals indicative of the incorporation or binding events can be detected to generate signal code sequences comprising a series of signal codes (corresponding to signals (ON signals), absence of signals (OFF signals), or a combination thereof) detected in the sequential cycles. Decoding of the identifier sequences based at least in part of the signal code sequences can be used to detect and locate the corresponding analytes.
Provided in some aspects are methods of patterning a surface in situ for producing an array on the surface, for example, by partitioning of beads comprising oligonucleotides into spatially predefined regions, to generate unique DNA sequences in spatial positions in the array. Compositions such as nucleic acid arrays produced by the methods are also disclosed.
C12Q 1/6806 - Préparation d’acides nucléiques pour analyse, p.ex. pour test de réaction en chaîne par polymérase [PCR]
B01L 3/00 - Récipients ou ustensiles pour laboratoires, p.ex. verrerie de laboratoire; Compte-gouttes
C12Q 1/6874 - Méthodes de séquençage faisant intervenir des réseaux d’acides nucléiques, p.ex. séquençage par hybridation [SBH]
C12Q 1/6888 - Produits d’acides nucléiques utilisés dans l’analyse d’acides nucléiques, p.ex. amorces ou sondes pour la détection ou l’identification d’organismes
23.
SINGLE CELL ANALYSIS OF TRANSPOSASE ACCESSIBLE CHROMATIN
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.
Provided herein are methods of improving sensitivity of spatial detection of an analyte in a biological sample using splint oligonucleotides, circularized second strands, and rolling circle amplification. For example, provided herein are methods of improving sensitivity of spatial detection of an analyte in a biological sample where a splint oligonucleotide hybridizes to a second strand; the second strand is ligated together thereby creating a circularized second strand, rolling circle amplification of the circularized second strand results in generation of an amplified second strand, and all or part of the sequence of the amplified second strand is determined and used to spatially detect the analyte in the biological sample.
Aspects of the present invention include analyzing nucleic acids from single cells using methods that include using tagged polynucleotides containing multiplex identifier sequences.
This disclosure relates to compositions and methods for three-dimensional spatial profiling of analytes in a biological sample. The methods include use of a hydrogel comprising one or more polymers that include a phenol moiety, an azide moiety, or an alkyne moiety.
A method of analyzing single biological particles, such as cells or nuclei, that maintains the single biological particles in a state of relative isolation during decrosslinking and subsequent processing is provided. In the case of cells, the method prevents cellular analytes from each individual cell from leaving the cell site and diffusing toward adjacent cells, while permitting transmission of a decrosslinking agent, followed by processing of the cellular analytes from the individual cells by barcoding and/or imaging of the cellular analytes.
C12Q 1/68 - Procédés de mesure ou de test faisant intervenir des enzymes, des acides nucléiques ou des micro-organismes; Compositions à cet effet; Procédés pour préparer ces compositions faisant intervenir des acides nucléiques
C12Q 1/6804 - Analyse d’acides nucléiques utilisant des immunogènes
C12Q 1/6806 - Préparation d’acides nucléiques pour analyse, p.ex. pour test de réaction en chaîne par polymérase [PCR]
Provided herein are methods of determining a location of an analyte in a biological sample, and systems for performing these methods. In particular, the methods and systems comprise decreasing the binding events of an analyte from a biological sample to capture probes that surround a biological sample positioned on a substrate.
The present disclosure relates in some aspects to methods for manufacturing molecular arrays using a hybrid approach comprising microfluidics-based delivery and photolithography-guided oligonucleotide hybridization and ligation. In particular, the molecular arrays can be used for determining spatial patterns of abundance and/or expression of a biological target in a sample.
The present disclosure relates in some aspects to methods and compositions for processes for inverting oligonucleotide molecules in an in situ synthesized array, including reversing the orientation of the oligonucleotide molecules with respect to the array substrate from 3′-immobilized to 5′-immobilized.
Provided in some aspects are methods for light-controlled in situ surface patterning of a substrate comprising a step of blocking or ablating oligonucleotide molecules in a boundary region separating a plurality of spot regions, and attaching oligonucleotides to oligonucleotide molecules in a first one or more of the spot regions.
G03F 7/11 - Matériaux photosensibles - caractérisés par des détails de structure, p.ex. supports, couches auxiliaires avec des couches de recouvrement ou des couches intermédiaires, p.ex. couches d'ancrage
The present disclosure relates generally to compositions, methods, kits, partitions and systems for high-throughput microfluidic identification of antigen-binding molecules and antigen binding molecule epitope mapping. Particularly, the compositions, methods, kits, partitions and systems of the disclosure relate to the characterization of cell-expressed antigen-binding molecules (e.g., antibodies) produced by a population of cells, such as B cells, that may have different binding and epitope specificity to a target antigen of interest and/or with enhanced activity.
The present disclosure relates generally to compositions, methods, and systems for the characterization of antigen-binding molecules (e.g., antibodies) in biological samples. This characterization permits the identification of the ABM that bind to regions of interest, or the mapping of ABMs according to their binding to specific regions of interest, of an antigen. Understanding the binding characteristics of ABMs at a region of interest level may facilitate the identification and production of immunotherapeutic molecules having desired properties.
C12Q 1/6804 - Analyse d’acides nucléiques utilisant des immunogènes
G01N 33/50 - Analyse chimique de matériau biologique, p.ex. de sang ou d'urine; Test par des méthodes faisant intervenir la formation de liaisons biospécifiques par ligands; Test immunologique
The present disclosure provides methods, compositions and systems for analyzing individual cells or cell populations through a partitioned analysis of contents of individual cells or cell populations, such as cancer cells and cells of the immune system. Individual cells or cell populations may be co-partitioned with processing reagents for accessing cellular contents, and for uniquely identifying the content of a given cell or cell population, and subsequently analyzing the content of the cell and characterizing it as having derived from an individual cell or cell population, including analysis and characterization of nucleic acid(s) from the cell through sequencing.
The present disclosure in some aspects relates to methods and compositions for improved detection and quantification of one or more analytes present in a biological sample. In some aspects, the methods and compositions provided herein address issues associated with the heterogeneity of rolling circle amplification (RCA) products during in situ analyses. In some aspects, the methods and compositions disclosed herein provide more homogenous RCA products for improved sample imaging.
Crosslinked fluorocarbon surface coating, kits therefor, microfluidic devices having crosslinked fluoropolymer surface coatings, and methods of use for droplet generation are provided. The kits, devices, and their methods have a droplet source region that is coated with a crosslinked fluoropolymer surface coating produced by the reaction of a fluorocarbon silane and a fluorocarbon polyol.
C08G 65/00 - Composés macromoléculaires obtenus par des réactions créant une liaison éther dans la chaîne principale de la macromolécule
C08G 65/336 - Polymères modifiés par post-traitement chimique avec des composés organiques contenant du silicium
C09D 171/00 - Compositions de revêtement à base de polyéthers obtenus par des réactions créant une liaison éther dans la chaîne principale; Compositions de revêtement à base de dérivés de tels polymères
40.
METHODS AND SYSTEMS FOR LIGHT-CONTROLLED SURFACE PATTERNING USING PHOTOMASKS
Provided in some aspects are methods for light-controlled in situ surface patterning of a substrate using a minimal mask scheme. Systems and kits employing the methods are also disclosed. In some embodiments, a method disclosed herein comprises using photomasks and photoresist for photocontrollable hybridization and/or ligation of nucleic acid molecules, wherein photoresist removal allows hybridization and/or ligation of nucleic acid molecules at the exposed area. A large diversity of barcodes can be created in molecules on the substrate via sequential rounds of light exposure, hybridization, and ligation.
C12Q 1/6874 - Méthodes de séquençage faisant intervenir des réseaux d’acides nucléiques, p.ex. séquençage par hybridation [SBH]
C12Q 1/6876 - Produits d’acides nucléiques utilisés dans l’analyse d’acides nucléiques, p.ex. amorces ou sondes
G03F 7/00 - Production par voie photomécanique, p.ex. photolithographique, de surfaces texturées, p.ex. surfaces imprimées; Matériaux à cet effet, p.ex. comportant des photoréserves; Appareillages spécialement adaptés à cet effet
41.
METHODS AND SYSTEMS FOR CHARACTERIZING ANALYTES FROM INDIVIDUAL CELLS OR CELL POPULATIONS
The present disclosure provides methods of processing or analyzing a sample. A method for processing a sample may comprise hybridizing a probe molecule to a target region of a nucleic acid molecule (e.g., a ribonucleic acid (RNA) molecule), barcoding the probe-nucleic acid molecule complex, and performing extension, denaturation, and amplification processes. A method for processing a sample may comprise hybridizing first and second probes to adjacent or non-adjacent target regions of a nucleic acid molecule (e.g., an RNA molecule), linking the first and second probes to provide a probe-linked nucleic acid molecule, and barcoding the probe-linked nucleic acid molecule. One or more processes of the methods described herein may be performed within a partition, such as a droplet or well. One or more processes of the methods described herein may be performed on a cell, such as a permeabilized cell.
C12N 15/10 - Procédés pour l'isolement, la préparation ou la purification d'ADN ou d'ARN
C12N 15/11 - Fragments d'ADN ou d'ARN; Leurs formes modifiées
C12N 15/85 - Vecteurs ou systèmes d'expression spécialement adaptés aux hôtes eucaryotes pour cellules animales
C12Q 1/6804 - Analyse d’acides nucléiques utilisant des immunogènes
C12Q 1/6806 - Préparation d’acides nucléiques pour analyse, p.ex. pour test de réaction en chaîne par polymérase [PCR]
C12Q 1/6818 - Tests d’hybridation caractérisés par les moyens de détection impliquant l’interaction de plusieurs marqueurs, p.ex. transfert d’énergie de résonance
C12Q 1/6827 - Tests d’hybridation pour la détection de mutation ou de polymorphisme
C12Q 1/6881 - Produits d’acides nucléiques utilisés dans l’analyse d’acides nucléiques, p.ex. amorces ou sondes pour le typage de tissu ou de cellule, p.ex. sondes d’antigène leucocytaire humain [HLA]
G01N 33/50 - Analyse chimique de matériau biologique, p.ex. de sang ou d'urine; Test par des méthodes faisant intervenir la formation de liaisons biospécifiques par ligands; Test immunologique
G01N 33/53 - Tests immunologiques; Tests faisant intervenir la formation de liaisons biospécifiques; Matériaux à cet effet
G01N 33/543 - Tests immunologiques; Tests faisant intervenir la formation de liaisons biospécifiques; Matériaux à cet effet avec un support insoluble pour l'immobilisation de composés immunochimiques
G01N 33/569 - Tests immunologiques; Tests faisant intervenir la formation de liaisons biospécifiques; Matériaux à cet effet pour micro-organismes, p.ex. protozoaires, bactéries, virus
G01N 33/58 - Analyse chimique de matériau biologique, p.ex. de sang ou d'urine; Test par des méthodes faisant intervenir la formation de liaisons biospécifiques par ligands; Test immunologique faisant intervenir des substances marquées
The present disclosure relates in some aspects to recombinant Bacillus phage AP50 polymerases, modified AP50 polymerases, and compositions thereof. Also provided are methods of using the recombinant and/or modified AP50 polymerases for nucleic acid amplification (e.g., rolling circle amplification). In some aspects, the compositions and methods disclosed herein provide more robust amplification (e.g., RCA) reactions for improved in vitro and in situ analysis.
Provided herein are methods for preparing biological samples for spatial proteomic analysis, methods of determining a location of a protein analyte in a biological sample, and methods of determining a location of a protein analyte and a nucleic acid analyte in a biological sample.
G01N 33/68 - Analyse chimique de matériau biologique, p.ex. de sang ou d'urine; Test par des méthodes faisant intervenir la formation de liaisons biospécifiques par ligands; Test immunologique faisant intervenir des protéines, peptides ou amino-acides
C12N 15/10 - Procédés pour l'isolement, la préparation ou la purification d'ADN ou d'ARN
C12Q 1/6804 - Analyse d’acides nucléiques utilisant des immunogènes
C12Q 1/6874 - Méthodes de séquençage faisant intervenir des réseaux d’acides nucléiques, p.ex. séquençage par hybridation [SBH]
G01N 33/58 - Analyse chimique de matériau biologique, p.ex. de sang ou d'urine; Test par des méthodes faisant intervenir la formation de liaisons biospécifiques par ligands; Test immunologique faisant intervenir des substances marquées
Provided herein are methods, compositions, and kits for reducing capture of analytes from a biological sample on a spatial array on areas not covered by a biological sample.
The present disclosure provides methods and systems for nucleic acid preparation and/or analysis. Nucleic acids may be derived from one or more cells. Nucleic acid preparation may comprise generating nucleic acid molecules of varying lengths. Nucleic acid analysis may comprise identifying nucleic acid sequence information with nucleosome position information.
Systems and methods for analyzing a tissue sample are provided. A set of images of the tissue on a substrate comprising capture spots is obtained. Each image is an emission image of the tissue upon excitation of the tissue at a corresponding excitation wavelength for a respective detectable marker. A first spatial dataset is acquired including, for each detectable marker for each respective image, a measured intensity of each capture spot indexed by a representative spatial barcode in a plurality of spatial barcodes. A second spatial dataset is acquired including nucleic acid quantification data representing a number of nucleic acid molecules originating from the tissue on the substrate and localized to each capture spot by an associated spatial barcode. At least a portion of the first and second spatial datasets co-registered to each other by the spatial barcodes are spatially displayed, on a display.
The present disclosure generally relates to compositions and methods for secreted analyte detection and analysis. The analytes detected and analyzed are mapped to a cell or nucleus having secreted the analyte and are useful to characterize the cell or nucleus. The ability to understand analyte secretion events has implications for the improvement of drug delivery, cell profiling, and diagnostic development.
G01N 33/569 - Tests immunologiques; Tests faisant intervenir la formation de liaisons biospécifiques; Matériaux à cet effet pour micro-organismes, p.ex. protozoaires, bactéries, virus
G01N 33/53 - Tests immunologiques; Tests faisant intervenir la formation de liaisons biospécifiques; Matériaux à cet effet
G01N 33/68 - Analyse chimique de matériau biologique, p.ex. de sang ou d'urine; Test par des méthodes faisant intervenir la formation de liaisons biospécifiques par ligands; Test immunologique faisant intervenir des protéines, peptides ou amino-acides
A method for colocalization obtains a first image encoding spatial abundance data for a nucleic acid from a biological sample through pixel values of subset of pixels in the first image. A second image is obtained encoding spatial abundance data for a non-nucleic acid analyte from the sample through pixel intensity values of subset of pixels in the second image. Thresholds of the first and second image are obtained. A determination, using a registration between the first and second images, of a first summation of the value of each pixel in the subset of pixels of the first image in which the corresponding pixel in the second image exceeds the second threshold value is made. The first summation, normalized to a summation of each value of each pixel in the subset of pixels of the first image, provides a measure of colocalization between the nucleic acid and non-nucleic acid analyte.
G16B 15/00 - TIC spécialement adaptées à l’analyse de structures moléculaires bidimensionnelles ou tridimensionnelles, p.ex. relations structurelles ou fonctionnelles ou alignement de structures
G16B 25/10 - Profilage de l’expression de gènes ou de protéines; Estimation ou normalisation de ratio d’expression
The present disclosure relates in some aspects to recombinant polymerases, variant PumA1 polymerases, and compositions thereof. Also provided herein are methods of using the recombinant polymerases and/or variant PumA1 polymerases for nucleic acid amplification (e.g., rolling circle amplification). In some aspects, the compositions and methods disclosed herein provide more robust amplification (e.g., RCA) reactions for improved in vitro and in situ analysis.
Methods and systems for selecting a cell of interest based on immune cell data are disclosed. For example, a method may comprise obtaining a single cell or spatial dataset, wherein the single cell or spatial dataset comprises a dataset of immune cell receptors, antibodies, or fragments thereof from a sample; identifying a clonotype group in the single cell or spatial dataset;
Methods and systems for selecting a cell of interest based on immune cell data are disclosed. For example, a method may comprise obtaining a single cell or spatial dataset, wherein the single cell or spatial dataset comprises a dataset of immune cell receptors, antibodies, or fragments thereof from a sample; identifying a clonotype group in the single cell or spatial dataset;
selecting a schema to visualize selected amino acids in the clonotype group based on positions or chemical identity of the selected amino acids; visualizing the selected amino acids in the clonotype group in a graphic representation according to the schema; and selecting a cell of interest from the clonotype group based on a pre-defined criterion using the graphic representation.
G01N 33/50 - Analyse chimique de matériau biologique, p.ex. de sang ou d'urine; Test par des méthodes faisant intervenir la formation de liaisons biospécifiques par ligands; Test immunologique
G01N 33/563 - Tests immunologiques; Tests faisant intervenir la formation de liaisons biospécifiques; Matériaux à cet effet faisant intervenir des fragments d'anticorps
53.
COMPOSITIONS AND METHODS OF MAKING GENE EXPRESSION LIBRARIES
The present disclosure provides systems and methods for processing or analyzing a sample. Methods may include generating supports (e.g., beads) comprising barcode molecules coupled thereto. A support comprising barcode molecules may be useful for analyzing or processing one or more analytes such as nucleic acid molecules, proteins, and/or perturbation agents.
C12N 15/10 - Procédés pour l'isolement, la préparation ou la purification d'ADN ou d'ARN
C12N 15/11 - Fragments d'ADN ou d'ARN; Leurs formes modifiées
C12N 15/85 - Vecteurs ou systèmes d'expression spécialement adaptés aux hôtes eucaryotes pour cellules animales
C12Q 1/6804 - Analyse d’acides nucléiques utilisant des immunogènes
C12Q 1/6806 - Préparation d’acides nucléiques pour analyse, p.ex. pour test de réaction en chaîne par polymérase [PCR]
C12Q 1/6818 - Tests d’hybridation caractérisés par les moyens de détection impliquant l’interaction de plusieurs marqueurs, p.ex. transfert d’énergie de résonance
C12Q 1/6827 - Tests d’hybridation pour la détection de mutation ou de polymorphisme
C12Q 1/6881 - Produits d’acides nucléiques utilisés dans l’analyse d’acides nucléiques, p.ex. amorces ou sondes pour le typage de tissu ou de cellule, p.ex. sondes d’antigène leucocytaire humain [HLA]
G01N 33/50 - Analyse chimique de matériau biologique, p.ex. de sang ou d'urine; Test par des méthodes faisant intervenir la formation de liaisons biospécifiques par ligands; Test immunologique
G01N 33/53 - Tests immunologiques; Tests faisant intervenir la formation de liaisons biospécifiques; Matériaux à cet effet
G01N 33/543 - Tests immunologiques; Tests faisant intervenir la formation de liaisons biospécifiques; Matériaux à cet effet avec un support insoluble pour l'immobilisation de composés immunochimiques
G01N 33/569 - Tests immunologiques; Tests faisant intervenir la formation de liaisons biospécifiques; Matériaux à cet effet pour micro-organismes, p.ex. protozoaires, bactéries, virus
G01N 33/58 - Analyse chimique de matériau biologique, p.ex. de sang ou d'urine; Test par des méthodes faisant intervenir la formation de liaisons biospécifiques par ligands; Test immunologique faisant intervenir des substances marquées
55.
METHODS, SYSTEMS, AND DEVICES FOR SAMPLE INTERFACE
An assembly includes a stage body, a cam plate disposed on the body, and a sample positioning plate having a sample positioning surface configured to receive a sample device. The surface has first, second, and third raised portions. The second raised portion is disposed between the first and third raised portions. The first, second, and third raised portions are configured to contact the sample device. The assembly includes a riser module disposed within the stage body. The riser module is coupled to the plate and the body. The assembly includes a lid configured to be coupled to the cam plate. The lid has a coupled configuration and an uncoupled configuration such that, when in the coupled configuration, a recess is formed by the lid and the plate. The assembly includes one or more light sources disposed in the cam plate that are configured to direct light within the recess.
The present disclosure relates in some aspects to methods and compositions for analyzing a target nucleic acid in a biological sample. In some aspects, the methods disclosed herein promote hybridization of polynucleotides to target nucleic acids and/or splints for enhanced ligation efficiency and/or enhanced hybridization efficiency in situ in a biological sample. In some aspects, the presence, amount, and/or identity of a target nucleic acid is analyzed in situ. Also provided are compositions and kits for use in accordance with the methods.
The present disclosure generally relates to methods and compositions for in situ analysis or detection of analytes in a biological sample. More specifically, the present disclosure relates to methods for reducing autofluorescence in tissue samples, methods for analyzing tissue samples, and compounds for use in the same. The methods and compounds of the present disclosure may be especially suitable for analytical methods employing fluorescence in situ hybridization techniques over multiple cycles of imaging.
Provided herein are methods, compositions, and kits for determining a location of a target analyte in a biological sample that include the use of terminal deoxynucleotidyl transferase.
Systems and methods for analyzing a tissue sample are provided. A set of images of the tissue on a substrate comprising capture spots is obtained. Each image is an emission image of the tissue upon excitation of the tissue at a corresponding excitation wavelength for a respective detectable marker. A first spatial dataset is acquired including, for each detectable marker for each respective image, a measured intensity of each capture spot indexed by a representative spatial barcode in a plurality of spatial barcodes. A second spatial dataset is acquired including nucleic acid quantification data representing a number of nucleic acid molecules originating from the tissue on the substrate and localized to each capture spot by an associated spatial barcode. At least a portion of the first and second spatial datasets co-registered to each other by the spatial barcodes are spatially displayed, on a display.
Various embodiments of the present disclosure disclose a system for forming a sealed chamber for preparing samples therein. In particular, various embodiments present an assembly that includes a sample device and a lid for forming a sealed chamber that can be used for, among other things, incubating samples therein. When the lid engages with the sample device, a plurality of snap joint elements of the lid engage with a respective plurality of snap joint elements to form a seal between the lid and the sample device.
A sample holder is provided. The sample holder includes a first member including a first retaining mechanism configured to retain a first substrate comprising a sample. The first retaining mechanism configured to retain the first substrate disposed in a first plane. The sample holder further includes a second member including a second retaining mechanism configured to retain a second substrate including a reagent medium disposed in a second plane. The sample holder further includes an alignment mechanism connected to one or both of the first member and the second member. The alignment mechanism configured to align the first and second members along the first plane and/or the second plane such that the sample contacts at least a portion of the reagent medium when the first and second members are aligned and within a threshold distance along an axis orthogonal to the second plane.
The present disclosure relates in some aspects to methods and compositions for in situ analysis involving catalytic de-crosslinking of biological samples.
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.
A substrate holder includes a base configured to receive a substrate; a cover configured to mateably engage with the base, the cover defining an opening formed by inner sidewalls; and a removable insert defining a surface, the removable insert being configured to be received within the opening of the cover. The removable insert includes a gasket; a projection coupled to the gasket; and at least two insert tabs extending from opposite sides of the removable insert, each insert tab being configured to engage with at least one of the inner sidewalls forming the opening of the cover.
The present disclosure generally relates to methods and compositions for detecting a plurality of molecules of one or more analytes in a sample. In some aspects, the present disclosure relates to methods for determining the locations and identities of analytes in a biological sample using detectably labeled probes comprising detectable labels with different signal emission lifetimes. In some aspects, the present disclosure relates to methods for identifying the detectable labels of the detectably labeled probes using time-gated detection. The methods herein have particular applicability in the detection of identifier sequences (e.g., analyte sequences or barcode sequences) in situ in a biological sample, including those using sequential cycles of detectably labeled probe hybridization to decode the identifier sequences.
The present disclosure generally relates to methods and compositions for in situ analysis or detection of analytes in a sample. More specifically, the present disclosure relates to methods for reducing autofluorescence in tissue samples, methods for analyzing biological samples, and compounds for use in the same. The methods and compounds of the present disclosure may be especially suitable for analytical methods employing fluorescence in situ hybridization techniques over multiple cycles of imaging.
The present disclosure relates in some aspects to methods and compositions for in situ analysis using nucleic acid complexes comprising a detectable label and a moiety that can be activated to extinguish signals of the detectable label. In some embodiments, the activatable moiety comprises a photosensitizer. The nucleic acid complexes may allow for improved detection and decreased signal carryover between detection cycles.
G01N 21/35 - Couleur; Propriétés spectrales, c. à d. comparaison de l'effet du matériau sur la lumière pour plusieurs longueurs d'ondes ou plusieurs bandes de longueurs d'ondes différentes en recherchant l'effet relatif du matériau pour les longueurs d'ondes caractéristiques d'éléments ou de molécules spécifiques, p.ex. spectrométrie d'absorption atomique en utilisant la lumière infrarouge
The present disclosure relates to methods, compositions, systems, and kits for detecting and analyzing the glycosylation of healthy and diseased cells and protein-specific glycosylation patterns using single-cell profiling methodologies.
G01N 33/68 - Analyse chimique de matériau biologique, p.ex. de sang ou d'urine; Test par des méthodes faisant intervenir la formation de liaisons biospécifiques par ligands; Test immunologique faisant intervenir des protéines, peptides ou amino-acides
G01N 33/53 - Tests immunologiques; Tests faisant intervenir la formation de liaisons biospécifiques; Matériaux à cet effet
In some aspects, the present disclosure relates to methods for reducing the detection of false positive events during analysis of a biological sample. In some aspects, the method comprises use of a decoy oligonucleotide that can hybridize to a probe or to a target nucleic acid. The methods herein have particular applicability in reducing the detection of false positive events and in combining hybridization and ligation reactions into a single step. Also provided are kits comprising probes for use in such methods.
The present disclosure relates in some aspects to methods and compositions for analyzing biological samples involving active stripping of detectably labeled probes from hybridization complexes such as rolling circle amplification products (RCPs). In some embodiments, the present application provides a method wherein double-stranded hybridized complexes are dissociated by DNA helicases. In some embodiments, single-stranded binding proteins facilitate the dissociation and prevent hybridization of the dissociated strands.
The present disclosure relates in some aspects to methods for analyzing antigen receptor transcripts in a biological sample. In some aspects, nucleic acid molecules are generated from V(D)J transcripts in situ in the biological sample to enrich molecules comprising V(D)J joins. In some aspects, the presence, amount, and/or identity of a plurality of V(D)J transcripts are analyzed in situ. Also provided are oligonucleotides, sets of oligonucleotides, compositions, and kits for use in accordance with the methods.
The present disclosure relates in some aspects to methods and compositions for manufacturing molecular arrays using a hybrid approach comprising using non-contact printing (e.g., using inkjet printing, slot die coating, and/or blade coating) and photolithography-guided oligonucleotide hybridization and ligation. In particular, the molecular arrays can be used for determining spatial patterns of abundance and/or expression of a biological target in a sample.
Provided herein are systems and methods for processing biomolecules (e.g., nucleic acid molecules, proteins) from a sample. A method for processing biomolecules may comprise hybridizing a probe molecule to a target region of a nucleic acid molecule (e.g., a ribonucleic acid (RNA) molecule) and barcoding the probe-nucleic acid molecule complex or derivatives thereof. Such a method can comprise performing a nucleic acid reaction, e.g., extension, denaturation, and amplification. A method for processing a sample may comprise hybridizing probes to (i) target regions of a nucleic acid molecule (e.g., RNA molecule) and (ii) a reporter oligonucleotide of a feature binding group, and barcoding the probe-associated molecules. One or more processes of the methods described herein may be performed within a partition, such as a droplet or well.
C12N 15/10 - Procédés pour l'isolement, la préparation ou la purification d'ADN ou d'ARN
C12N 15/11 - Fragments d'ADN ou d'ARN; Leurs formes modifiées
C12N 15/85 - Vecteurs ou systèmes d'expression spécialement adaptés aux hôtes eucaryotes pour cellules animales
C12Q 1/6804 - Analyse d’acides nucléiques utilisant des immunogènes
C12Q 1/6806 - Préparation d’acides nucléiques pour analyse, p.ex. pour test de réaction en chaîne par polymérase [PCR]
C12Q 1/6818 - Tests d’hybridation caractérisés par les moyens de détection impliquant l’interaction de plusieurs marqueurs, p.ex. transfert d’énergie de résonance
C12Q 1/6827 - Tests d’hybridation pour la détection de mutation ou de polymorphisme
C12Q 1/6881 - Produits d’acides nucléiques utilisés dans l’analyse d’acides nucléiques, p.ex. amorces ou sondes pour le typage de tissu ou de cellule, p.ex. sondes d’antigène leucocytaire humain [HLA]
G01N 33/50 - Analyse chimique de matériau biologique, p.ex. de sang ou d'urine; Test par des méthodes faisant intervenir la formation de liaisons biospécifiques par ligands; Test immunologique
G01N 33/53 - Tests immunologiques; Tests faisant intervenir la formation de liaisons biospécifiques; Matériaux à cet effet
G01N 33/543 - Tests immunologiques; Tests faisant intervenir la formation de liaisons biospécifiques; Matériaux à cet effet avec un support insoluble pour l'immobilisation de composés immunochimiques
G01N 33/569 - Tests immunologiques; Tests faisant intervenir la formation de liaisons biospécifiques; Matériaux à cet effet pour micro-organismes, p.ex. protozoaires, bactéries, virus
G01N 33/58 - Analyse chimique de matériau biologique, p.ex. de sang ou d'urine; Test par des méthodes faisant intervenir la formation de liaisons biospécifiques par ligands; Test immunologique faisant intervenir des substances marquées
76.
DEVICES, METHODS, AND SYSTEMS FOR IMPROVED DROPLET RECOVERY
Devices, methods, and systems for generating droplets are provided. The devices, methods, and systems are designed to maximize droplet recovery, e.g., from a collection reservoir.
The present disclosure relates to methods, compositions, and kits for determining the location of target RNA in a biological sample and/or rescuing target RNA, from which spatial gene expression analysis is difficult to determine, from fresh-frozen biological samples.
C07K 16/28 - Immunoglobulines, p.ex. anticorps monoclonaux ou polyclonaux contre du matériel provenant d'animaux ou d'humains contre des récepteurs, des antigènes de surface cellulaire ou des déterminants de surface cellulaire
C12Q 1/00 - Procédés de mesure ou de test faisant intervenir des enzymes, des acides nucléiques ou des micro-organismes; Compositions à cet effet; Procédés pour préparer ces compositions
G01N 33/566 - Tests immunologiques; Tests faisant intervenir la formation de liaisons biospécifiques; Matériaux à cet effet utilisant un support spécifique ou des protéines réceptrices comme réactifs pour la formation de liaisons par ligand
80.
METHODS FOR SPATIAL ANALYSIS USING ROLLING CIRCLE AMPLIFICATION
Provided herein are methods of identifying a location of an analyte in a biological sample using padlock oligonucleotides and rolling circle amplification. For example, provided herein are methods for identifying a location of an analyte in a biological sample where (i) a padlock oligonucleotide hybridizes to an analyte and is ligated thereby creating a circularized padlock oligonucleotide, (ii) rolling circle amplification of the circularized padlock oligonucleotide results in generation of an amplified circularized padlock oligonucleotide, and (iii) a signal corresponding to the amplified circularized padlock oligonucleotide is detected and used to identify the location of the analyte in the biological sample. Also provided are methods for identifying whether a treatment of a biological sample facilitates the release and subsequent detection of an analyte from the biological sample using padlock oligonucleotides and rolling circle amplification.
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.
C12Q 1/6881 - Produits d’acides nucléiques utilisés dans l’analyse d’acides nucléiques, p.ex. amorces ou sondes pour le typage de tissu ou de cellule, p.ex. sondes d’antigène leucocytaire humain [HLA]
C12Q 1/6874 - Méthodes de séquençage faisant intervenir des réseaux d’acides nucléiques, p.ex. séquençage par hybridation [SBH]
C12Q 1/48 - Procédés de mesure ou de test faisant intervenir des enzymes, des acides nucléiques ou des micro-organismes; Compositions à cet effet; Procédés pour préparer ces compositions faisant intervenir une transférase
C12Q 1/6806 - Préparation d’acides nucléiques pour analyse, p.ex. pour test de réaction en chaîne par polymérase [PCR]
82.
METHODS AND SYSTEMS FOR CHARACTERIZING ANTIGEN-BINDING MOLECULES EXPRESSED BY IMMUNE CELLS
The present disclosure relates generally to methods and systems for the characterization of antigen-binding molecules, e.g., B cell receptors, antibodies, and antigen-binding fragments thereof, using single-cell immune profiling methodologies. The methods and systems described herein permit rapid, high-throughput identification and characterization of antigen binding molecules having desired properties.
G01N 33/569 - Tests immunologiques; Tests faisant intervenir la formation de liaisons biospécifiques; Matériaux à cet effet pour micro-organismes, p.ex. protozoaires, bactéries, virus
C12Q 1/68 - Procédés de mesure ou de test faisant intervenir des enzymes, des acides nucléiques ou des micro-organismes; Compositions à cet effet; Procédés pour préparer ces compositions faisant intervenir des acides nucléiques
C12N 15/10 - Procédés pour l'isolement, la préparation ou la purification d'ADN ou d'ARN
83.
COMPOSITIONS AND METHODS FOR CHARACTERIZING BINDING CHARACTERISTICS OF ANTIGEN BINDING MOLECULES FROM SINGLE CELLS
The present disclosure relates generally to compositions, methods, and systems for the characterization of binding charactersitics of antigen binding molecules, e.g., antibodies, using single-cell immune profiling methodologies. The compositions, methods and systems described herein permit rapid, high-throughput identification and characterization of antigen binding molecules having desired properties.
G01N 33/543 - Tests immunologiques; Tests faisant intervenir la formation de liaisons biospécifiques; Matériaux à cet effet avec un support insoluble pour l'immobilisation de composés immunochimiques
84.
METHODS FOR CHARACTERIZATION OF ANTIGEN-BINDING MOLECULES FROM BIOLOGICAL SAMPLES
C07K 16/00 - Immunoglobulines, p.ex. anticorps monoclonaux ou polyclonaux
C07K 16/30 - Immunoglobulines, p.ex. anticorps monoclonaux ou polyclonaux contre du matériel provenant d'animaux ou d'humains contre des récepteurs, des antigènes de surface cellulaire ou des déterminants de surface cellulaire provenant de cellules de tumeurs
The present disclosure provides compositions, methods, systems, and devices for polynucleotide processing. Such polynucleotide processing may be useful for a variety of applications, including polynucleotide sequencing.
C12Q 1/6874 - Méthodes de séquençage faisant intervenir des réseaux d’acides nucléiques, p.ex. séquençage par hybridation [SBH]
C12Q 1/6806 - Préparation d’acides nucléiques pour analyse, p.ex. pour test de réaction en chaîne par polymérase [PCR]
C12Q 1/683 - Tests d’hybridation pour la détection de mutation ou de polymorphisme faisant intervenir des enzymes de restriction, p.ex. polymorphisme de longueur de fragment de resctriction
C12Q 1/6804 - Analyse d’acides nucléiques utilisant des immunogènes
86.
MULTIPLEX CAPTURE OF GENE AND PROTEIN EXPRESSION FROM A BIOLOGICAL SAMPLE
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.
44 - Services médicaux, services vétérinaires, soins d'hygiène et de beauté; services d'agriculture, d'horticulture et de sylviculture.
Produits et services
Medical equipment, namely, devices for analyzing nucleic acids, for RNA and DNA analysis, and for DNA sequencing for the purpose of rendering medical treatment and diagnosis, and software for use therewith sold as a unit. Medical diagnostic testing and reporting services in the fields of DNA sequencing, RNA and DNA analysis, and genetic analysis.
88.
METHODS AND SYSTEMS FOR PROCESSING POLYNUCLEOTIDES
This disclosure provides methods and compositions for sample processing, particularly for sequencing applications. Included within this disclosure are bead compositions, such as diverse libraries of beads attached to large numbers of oligonucleotides containing barcodes. Often, the beads provides herein are degradable. For example, they may contain disulfide bonds that are susceptible to reducing agents. The methods provided herein include methods of making libraries of barcoded beads as well as methods of combining the beads with a sample, such as by using a microfluidic device.
The present disclosure provides methods and systems for cell and bead processing or analysis. A method for processing a cell or bead may comprise subjecting a bead to conditions sufficient to change a first characteristic or set of characteristics (e.g., cell or bead size). Such a method may further comprise subjecting the cell or bead to conditions sufficient to change a second characteristic or set of characteristics. In some cases, crosslinks may be formed within the cell or bead.
C12N 5/00 - Cellules non différenciées humaines, animales ou végétales, p.ex. lignées cellulaires; Tissus; Leur culture ou conservation; Milieux de culture à cet effet
90.
ANALYSIS OF ANTIGEN AND ANTIGEN RECEPTOR INTERACTIONS
The present disclosure relates in some aspects to methods of contacting a biological sample (e.g., fresh or frozen tissue sample) with peptide-loaded detection complexes. In some aspects, the peptide-loaded detection complexes comprise antigen presenting molecule monomers or multimers that bind to antigens, reporter oligonucleotides that correspond to the antigen/antigen presenting molecule combination. In some aspects, the peptide-loaded detection complexes are tetramers. In some aspects, the tetramers bound to T cell receptors of T cells may be detected in situ. In some aspects, the tetramers bound to TCRs may be detected on an in situ platform and/or using an array comprising spatially barcoded capture agents. In some embodiments, the method comprises contacting a plurality of peptide-loaded detection complexes to clonal populations of T cells comprising TCRs of different antigen specificities.
Disclosed herein are methods of amplifying an analyte in a biological sample using a bridging oligonucleotide that hybridizes to a captured analyte. The methods disclosed herein include steps of (a) contacting a biological sample with a substrate having capture probes comprising a capture domain and a spatial barcode; (b) hybridizing the analyte to the capture domain; and (c) contacting the analyte to a bridging oligonucleotide comprising (i) a capture-probe-binding sequence, and (ii) an analyte-binding sequence; (d) extending the bridging oligonucleotide; and (e) determining (i) all or a part of the sequence of the analyte, or a complement thereof, and (ii) the spatial barcode, or a complement thereof, and using the determined sequence of (i) and (ii) to determine the location of the analyte in the biological sample.
The present disclosure relates in some aspects to methods and compositions for light-controlled in situ surface patterning of a substrate using a Click chemistry reaction. In some embodiments, a method disclosed herein comprises a copper-catalyzed alkyne-azide (CuAAC) click chemistry reaction for photocontrollable ligation of nucleic acid molecules. A large diversity of barcodes can be created in molecules on the substrate via sequential rounds of light exposure, hybridization, and ligation.
G01N 33/50 - Analyse chimique de matériau biologique, p.ex. de sang ou d'urine; Test par des méthodes faisant intervenir la formation de liaisons biospécifiques par ligands; Test immunologique
A61K 39/00 - Préparations médicinales contenant des antigènes ou des anticorps
The present disclosure relates in some aspects to methods for manufacturing molecular arrays using a hybrid approach comprising microfluidics-based delivery and photolithography- guided oligonucleotide hybridization and ligation. In particular, the molecular arrays can be used for determining spatial patterns of abundance and/or expression of a biological target in a sample.
The present disclosure relates in some aspects to methods and compositions for processes for inverting oligonucleotide molecules in an in situ synthesized array, including reversing the orientation of the oligonucleotide molecules with respect to the array substrate from 3'-immobilized to 5'-immobilized using an oligonucleotide immobilized on the substrate via its 5' terminus. The method may comprise using a primer sequence for extension and/or a splint for ligation.
C12Q 1/6837 - Couplage enzymatique ou biochimique d’acides nucléiques à une phase solide utilisant des réseaux de sondes ou des puces à sondes
C40B 50/14 - Synthèse en phase solide, c. à d. dans laquelle au moins un bloc servant à créer la bibliothèque est lié à un support solide au cours de la création de la bibliothèque; Procédés particuliers de clivage à partir du support solide
C40B 50/18 - Synthèse en phase solide, c. à d. dans laquelle au moins un bloc servant à créer la bibliothèque est lié à un support solide au cours de la création de la bibliothèque; Procédés particuliers de clivage à partir du support solide utilisant un procédé particulier d'ancrage au support solide
96.
COMPOSITIONS AND METHODS FOR GENERATING MOLECULAR ARRAYS USING OLIGONUCLEOTIDE PRINTING AND PHOTOLITHOGRAPHY
The present disclosure relates in some aspects to methods and compositions for manufacturing molecular arrays using a hybrid approach comprising using non-contact printing (e.g., using inkjet printing, slot die coating, and/or blade coating) and photolithography-guided oligonucleotide hybridization and ligation. In particular, the molecular arrays can be used for determining spatial patterns of abundance and/or expression of a biological target in a sample.
The present disclosure relates in some aspects to methods and compositions for light- controlled in situ surface patterning of a substrate using a Click chemistry reaction. In some embodiments, a method disclosed herein comprises a copper-catalyzed alkyne-azide (CuAAC) click chemistry reaction for photocontrollable ligation of nucleic acid molecules. A large diversity of barcodes can be created in molecules on the substrate via sequential rounds of light exposure, hybridization, and ligation.
Provided herein are reaction mixtures, compositions, systems, methods, and kits for assessing enzymatic activity. Aspects of the disclosure include use of a reactant barcoded oligonucleotide (RBO) construct comprising a first oligonucleotide comprising a reaction barcode sequence and a first linker that connects the first oligonucleotide to a first reactant, an amplification construct comprising a second oligonucleotide that is complementary to at least a portion of the first oligonucleotide and a second linker that connects the second oligonucleotide to a second reactant, and a blocking construct, comprising a third oligonucleotide that is complementary to at least a portion of the first oligonucleotide. A rate of reaction product generation may be measured to determine an enzymatic activity of a given enzyme.
C12Q 1/48 - Procédés de mesure ou de test faisant intervenir des enzymes, des acides nucléiques ou des micro-organismes; Compositions à cet effet; Procédés pour préparer ces compositions faisant intervenir une transférase
C12N 9/12 - Transférases (2.) transférant des groupes contenant du phosphore, p.ex. kinases (2.7)
99.
METHODS AND SYSTEMS FOR PROCESSING POLYNUCLEOTIDES
The present disclosure provides compositions, methods, systems, and devices for polynucleotide processing. Such polynucleotide processing may be useful for a variety of applications, including polynucleotide sequencing.
C12N 15/10 - Procédés pour l'isolement, la préparation ou la purification d'ADN ou d'ARN
C40B 20/04 - Identification des éléments d'une bibliothèque au moyen d'une étiquette, d'un marqueur ou d'un autre identificateur lisible ou détectable, p.ex. procédés de décodage
C12Q 1/6804 - Analyse d’acides nucléiques utilisant des immunogènes
C12Q 1/6806 - Préparation d’acides nucléiques pour analyse, p.ex. pour test de réaction en chaîne par polymérase [PCR]
C40B 50/16 - Synthèse en phase solide, c. à d. dans laquelle au moins un bloc servant à créer la bibliothèque est lié à un support solide au cours de la création de la bibliothèque; Procédés particuliers de clivage à partir du support solide comprenant des étapes de codage
C12Q 1/6874 - Méthodes de séquençage faisant intervenir des réseaux d’acides nucléiques, p.ex. séquençage par hybridation [SBH]
C12Q 1/683 - Tests d’hybridation pour la détection de mutation ou de polymorphisme faisant intervenir des enzymes de restriction, p.ex. polymorphisme de longueur de fragment de resctriction
100.
PROBE-BASED ANALYSIS OF NUCLEIC ACIDS AND PROTEINS
Provided herein are systems and methods for processing biomolecules (e.g., nucleic acid molecules, proteins) from a sample. A method for processing biomolecules may comprise hybridizing a probe molecule to a target region of a nucleic acid molecule (e.g., a ribonucleic acid (RNA) molecule) and barcoding the probe-nucleic acid molecule complex or derivatives thereof. Such a method can comprise performing a nucleic acid reaction, e.g., extension, denaturation, and amplification. A method for processing a sample may comprise hybridizing probes to (i) target regions of a nucleic acid molecule (e.g., RNA molecule) and (ii) a reporter oligonucleotide of a feature binding group, and barcoding the probe-associated molecules. One or more processes of the methods described herein may be performed within a partition, such as a droplet or well.
