公开(公告)号：US20240271328A1

公开(公告)日：2024-08-15

申请号：US18582106

申请日：2024-02-20

申请人： SEQONCE BIOSCIENCES, INC.

发明人： Joseph DUNHAM

IPC分类号： C40B50/06 ,  C12N15/10 ,  C40B70/00 ,  C40B80/00

CPC分类号： C40B50/06 ,  C12N15/1068 ,  C40B70/00 ,  C40B80/00

摘要： Rapid methods, capable of being performed in a single reaction tube, are described herein for constructing libraries for high-throughput polynucleotide sequencing applications, such as next generation sequencing (NGS) applications. Oligonucleotide probes include chemically-active groups at their 5′ or 3′ ends, or both, to facilitate the cleavage of their 5′ or 3′ ends, or both, following their hybridization to the single-stranded ends of frayed template fragments. Cleavage of probe ends reveal single-stranded regions at the ends of the hybridized fragments. Adaptors, specific to these ends, are ligated to the hybridized probe/template fragments, and blunt end fragments are ligated to blunt ends of hybridized probe/template fragments, if present, to generate the adaptor-ligated fragments of the library.

公开(公告)号：US12031176B2

公开(公告)日：2024-07-09

申请号：US16940517

申请日：2020-07-28

申请人： HITACHI HIGH-TECH CORPORATION

发明人： Masataka Shirai ,  Tomoyuki Sakai ,  Kenko Uchida

IPC分类号： C12Q1/68 ,  C12N15/10 ,  C12Q1/6806 ,  C12Q1/6809 ,  C12Q1/6837 ,  C40B40/08 ,  C40B50/06 ,  G01N33/543

CPC分类号： C12Q1/6837 ,  C12N15/1003 ,  C12N15/1093 ,  C12N15/1096 ,  C12Q1/68 ,  C12Q1/6806 ,  C12Q1/6809 ,  C40B40/08 ,  C40B50/06 ,  G01N33/54306

摘要： The purpose of the present invention is to provide a single cell analysis device in which the improvement of the nucleic acid capturing efficiency and the improvement of the cell capturing efficiency are both achieved and a highly accurate single cell analysis data is thereby obtained. The present invention relates to an improvement of a cell analysis device including a two-dimensional array chip having a plurality of cell capture parts capable of capturing a single cell in each of the capture parts, and nucleic acid capture parts corresponding to the respective cell capture parts, the nucleic acid capture parts being capable of capturing a nucleic acid extracted from the cell captured by the cell capture part.

公开(公告)号：US12024797B2

公开(公告)日：2024-07-02

申请号：US17133418

申请日：2020-12-23

申请人： GRAIL, LLC

发明人： Matthew H. Larson ,  Hyunsung John Kim ,  Nick Eattock ,  Xiao Yang

IPC分类号： C40B50/06 ,  C12N15/10 ,  C12Q1/6806 ,  C12Q1/6855 ,  C12Q1/6874

CPC分类号： C40B50/06 ,  C12N15/1065 ,  C12N15/1093 ,  C12Q1/6806 ,  C12Q1/6855 ,  C12Q1/6874 ,  C12Q1/6806 ,  C12Q2525/191 ,  C12Q2533/101 ,  C12Q2535/122 ,  C12Q2565/514 ,  C12N15/1093 ,  C12Q2525/191 ,  C12Q2533/101 ,  C12Q2535/122 ,  C12Q2563/179

摘要： Aspects of the invention relate to methods for preparing and analyzing a sequencing library from a mixed cell-free DNA (cfDNA) sample, wherein the mixed sample includes double-stranded DNA (dsDNA), damaged dsDNA (e.g., nicked dsDNA), and single-stranded DNA (ssDNA) molecules. The subject methods facilitate the collection of information from dsDNA, ssDNA and damaged DNA (e.g., nicked DNA) molecules in a sample, thereby providing enhanced diagnostic information as compared to sequencing libraries that are prepared from dsDNA alone.

公开(公告)号：US12018254B2

公开(公告)日：2024-06-25

申请号：US17386176

申请日：2021-07-27

申请人： Wisconsin Alumni Research Foundation

发明人： Srivatsan Raman ,  Phil Huss

IPC分类号： C40B50/06 ,  C12N9/22 ,  C12N15/10 ,  C12N15/11

CPC分类号： C12N15/1093 ,  C12N9/22 ,  C12N15/102 ,  C12N15/11 ,  C12N2310/20 ,  C12N2795/10052

摘要： Described herein is a method of preparing an unbiased library of phage variants, comprising (a) preparing a population of “acceptor phage”; (b) removing an endogenous target gene and inserting gene variants into the acceptor phage genomes; (c) enriching the recombined phages; and (d) expressing the library for selection. The acceptor phage is a lytic phage comprising a synthetic genome wherein the target gene of interest is flanked by recombinase sites. The acceptor phage infects a first host bacteria expressing a recombination plasmid facilitating recombination. The phages then infect a second host bacteria expressing a counterselection system that accumulates recombined phage variants and selecting against non-recombined phages. The accumulated phage variants infect a third host bacteria. The phage library may then be sequenced and characterized.

公开(公告)号：US12001962B2

公开(公告)日：2024-06-04

申请号：US18230273

申请日：2023-08-04

申请人： CATALOG TECHNOLOGIES, INC.

发明人： Nathaniel Roquet ,  Hyunjun Park ,  Swapnil P. Bhatia ,  Darren R. Link

IPC分类号： G06F16/00 ,  C12N9/22 ,  C12N15/10 ,  C40B50/06 ,  G06F16/22 ,  G06F16/245 ,  G06N3/123 ,  G11C13/00 ,  G16B30/00 ,  G16B30/20 ,  G16B50/00 ,  G16B99/00

CPC分类号： G06N3/123 ,  C12N9/22 ,  C12N15/1031 ,  C12N15/1089 ,  C40B50/06 ,  G06F16/2272 ,  G06F16/245 ,  G11C13/0019 ,  G16B30/00 ,  G16B30/20 ,  G16B50/00 ,  G16B99/00 ,  C12N2310/20

摘要： Methods and systems for encoding digital information in nucleic acid (e.g., deoxyribonucleic acid) molecules without base-by-base synthesis, by encoding bit-value information in the presence or absence of unique nucleic acid sequences within a pool, comprising specifying each bit location in a bit-stream with a unique nucleic sequence and specifying the bit value at that location by the presence or absence of the corresponding unique nucleic acid sequence in the pool. But, more generally, specifying unique bytes in a bytestream by unique subsets of nucleic acid sequences. Also disclosed are methods for generating unique nucleic acid sequences without base-by-base synthesis using combinatorial genomic strategies (e.g., assembly of multiple nucleic acid sequences or enzymatic-based editing of nucleic acid sequences).

公开(公告)号：US11978533B2

公开(公告)日：2024-05-07

申请号：US16607738

申请日：2017-04-26

申请人： HUNAN ZONSEN PEPLIB BIOTECH CO., LTD

发明人： Zhuying Wang ,  Xiangqun Li

IPC分类号： G16B35/10 ,  C12N15/10 ,  C40B50/06

CPC分类号： G16B35/10 ,  C12N15/1093 ,  C40B50/06

摘要： An improved peptide library preparation method is described for constructing complete virtual peptide libraries such as a complete virtual tripeptide library, tetrapeptide library, pentapeptide library, hexapeptide library, heptapeptide library, or a complete octapeptide library, etc. The method includes constructing an expression vector for the expression of cyclic peptides. Each cyclic peptide displays an array of peptides of different sizes and sequences, and the number of cyclic peptides needed for constructing a complete virtual peptide library can be dramatically reduced compared with conventional chemical peptide synthesis. Furthermore, the cyclic peptide libraries can be readily reproduced by the expression and purification of the cyclic peptides using the constructed gene libraries. The improved peptide library preparation method can particularly be used, for example, to construct a complete virtual tetrapeptide library, a complete virtual pentapeptide library, a complete virtual hexapeptide library, a complete virtual heptapeptide library, and so on. The improved peptide library preparation method can also be used, for example, to construct a partial pentapeptide library, a partial hexapeptide library, a partial heptapeptide library, and so on. Other related methods and the related expression vectors are also described.

公开(公告)号：US11912756B2

公开(公告)日：2024-02-27

申请号：US17236978

申请日：2021-04-21

申请人： NantBio, Inc.

发明人： Clifford Anders Olson

IPC分类号： C40B40/08 ,  C07K16/00 ,  C12N15/10 ,  C12N15/62 ,  C12N15/11 ,  C07K17/00 ,  C07K16/24 ,  C07K16/28 ,  C40B50/06 ,  C40B30/04

CPC分类号： C07K16/005 ,  C07K16/244 ,  C07K16/2827 ,  C07K17/00 ,  C12N15/1037 ,  C12N15/1041 ,  C12N15/1062 ,  C12N15/111 ,  C12N15/62 ,  C40B40/08 ,  C40B50/06 ,  C07K2317/515 ,  C07K2317/565 ,  C07K2317/622 ,  C07K2317/73 ,  C07K2317/92 ,  C07K2317/94 ,  C40B30/04

摘要： Compositions, methods and uses of high-diversity nucleic acid library that encodes a plurality of antibodies or antibody fragments are presented. The high-diversity nucleic acid library comprises or is derived from (1) a VH-CDR1/2 sub-library, (2) a plurality of VH-CDR3 sub-libraries, and (3) a VL sub-library, each of which comprises a plurality of members. Preferably, each member of the sub-libraries comprises at least one random cassette that has a plurality of degenerate base positions. In an especially preferred embodiment, at least portions of at least two members of the VH-CDR1/2 sub-library, the plurality of VH-CDR3 sub-libraries, and the VL sub-library are recombined to form an expression library member in an expression library, where each member of the expression library encodes a distinct antibody or antibody fragment.

公开(公告)号：US11898270B2

公开(公告)日：2024-02-13

申请号：US16628481

申请日：2017-08-21

申请人： HUAZHONG AGRICULTURAL UNIVERSITY

发明人： Shuhong Zhao ,  Shengsong Xie ,  Changzhi Zhao ,  Xinyun Li ,  Xiangdong Liu ,  Xiaosong Han ,  Gaojuan Yang ,  Yang Gao ,  Yilong Chen ,  Xiaoyong Du ,  Yiliang Miao ,  Yunlong Ma ,  Xiaolei Liu

IPC分类号： C12N15/90 ,  C40B40/02 ,  C12N9/22 ,  C12N15/113 ,  C12N15/85 ,  C12Q1/6869 ,  C40B40/06 ,  C40B50/06

CPC分类号： C40B40/02 ,  C12N9/22 ,  C12N15/113 ,  C12N15/85 ,  C12N15/907 ,  C12Q1/6869 ,  C40B40/06 ,  C40B50/06 ,  C12N2810/10

摘要： Provided is a pig genome-wide specific sgRNA library, a preparation method therefor, and an application thereof. The sgRNA is targeted at a pig genome-wide protein-coding gene, lincRNA and/or miRNA. Specifically, an sgRNA construct has the following structure: AL-N20-AR, wherein AL is the left homology arm sequence located at the upstream of the coding sequence of a pig specific SgRNA, N20 is the coding sequence of the pig specific SgRNA, and AR is a right homology arm sequence located at the downstream of the coding sequence of the pig specific sgRNA. The sgRNA library can be used for screening functional genes of a pig or for preparing a kit.

公开(公告)号：US11852628B2

公开(公告)日：2023-12-26

申请号：US18185215

申请日：2023-03-16

申请人： 10X GENOMICS, INC.

发明人： Tarjei Sigurd Mikkelsen ,  Eswar Prasad Ramachandran Iyer ,  Andrew Kohlway ,  Luigi Jhon Alvarado Martinez ,  Katherine Pfeiffer ,  Andrew D. Price

IPC分类号： G01N33/532 ,  C07K14/74 ,  C12N15/10 ,  C12N15/11 ,  C12N15/85 ,  C12Q1/6804 ,  C12Q1/6806 ,  C12Q1/6818 ,  C12Q1/6827 ,  C12Q1/6881 ,  G01N33/50 ,  G01N33/53 ,  G01N33/543 ,  G01N33/548 ,  G01N33/569 ,  G01N33/58 ,  C40B30/04 ,  C40B50/06 ,  C40B70/00

CPC分类号： G01N33/532 ,  C07K14/70539 ,  C12N15/1037 ,  C12N15/1055 ,  C12N15/1065 ,  C12N15/1075 ,  C12N15/11 ,  C12N15/85 ,  C12Q1/6804 ,  C12Q1/6806 ,  C12Q1/6818 ,  C12Q1/6827 ,  C12Q1/6881 ,  G01N33/505 ,  G01N33/5032 ,  G01N33/5304 ,  G01N33/5306 ,  G01N33/5308 ,  G01N33/548 ,  G01N33/54306 ,  G01N33/54366 ,  G01N33/56977 ,  G01N33/58 ,  C12N2310/20 ,  C12N2320/10 ,  C12Q2537/164 ,  C12Q2563/179 ,  C12Q2563/185 ,  C12Q2565/1015 ,  C40B30/04 ,  C40B50/06 ,  C40B70/00

摘要： 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.

公开(公告)号：US11807956B2

公开(公告)日：2023-11-07

申请号：US17068551

申请日：2020-10-12

申请人： Twist Bioscience Corporation

发明人： Anthony Cox ,  Sebastian Treusch ,  Siyuan Chen

IPC分类号： C40B50/06 ,  C12N15/10 ,  G01N33/50 ,  C40B50/14 ,  C40B50/08 ,  C12N15/113 ,  C12Q1/6876 ,  C12Q1/6886

CPC分类号： C40B50/06 ,  C12N15/1079 ,  C12N15/1093 ,  C12N15/113 ,  C12Q1/6876 ,  C40B50/08 ,  C40B50/14 ,  G01N33/502 ,  G01N33/5029 ,  G01N33/5041 ,  C12N2320/30 ,  C12N2330/31 ,  C12Q1/6886 ,  C12Q2600/158

摘要： Disclosed herein are methods for the generation of highly accurate oligonucleic acid libraries encoding for predetermined variants of a nucleic acid sequence. The degree of variation may be complete, resulting in a saturated variant library, or less than complete, resulting in a selective library of variants. The variant oligonucleic acid libraries described herein may designed for further processing by transcription or translation. The variant oligonucleic acid libraries described herein may be designed to generate variant RNA, DNA and/or protein populations. Further provided herein are method for identifying variant species with increased or decreased activities, with applications in regulating biological functions and the design of therapeutics for treatment or reduction of disease.