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432 条记录 (耗时 0.034 秒)

    GENE EXPRESSION ANALYSIS IN SINGLE CELLS
    42.
    发明公开
    GENE EXPRESSION ANALYSIS IN SINGLE CELLS 审中-公开
    EINZELZELLEN中的表达蛋白酶

    公开(公告)号：EP3002337A1

    公开(公告)日：2016-04-06

    申请号：EP15190426.5

    申请日：2010-03-23

    申请人： Illumina, Inc.

    发明人： LINNARSSON, Sten

    IPC分类号： C12Q1/68 ,  C12N15/11 ,  C40B50/06

    CPC分类号： C12N15/1065 ,  C12N15/1096

    摘要： The present invention provides methods and compositions for the analysis of gene expression in single cells or in a plurality of single cells. The invention provides methods for preparing a cDNA library from individual cells by releasing mRNA from each single cell to provide a plurality of individual mRNA samples, synthesizing cDNA from the individual mRNA samples, tagging the individual cDNA, pooling the tagged cDNA samples and amplifying the pooled cDNA samples to generate a cDNA library. The invention also provides a cDNA library produced by the methods described herein. The invention further provides methods for analyzing gene expression in a plurality of cells by preparing a cDNA library as described herein and sequencing the library.

    摘要翻译： 本发明提供用于分析单细胞或多个单细胞中基因表达的方法和组合物。 本发明提供了通过从每个单个细胞释放mRNA从单个细胞制备cDNA文库以提供多个单个mRNA样品，合成来自各个mRNA样品的cDNA，标记单个cDNA，汇集标记的cDNA样品并扩增合并的 cDNA样品以产生cDNA文库。 本发明还提供了通过本文所述方法产生的cDNA文库。 本发明进一步提供了通过制备本文所述的cDNA文库并测序文库来分析多个细胞中的基因表达的方法。

    METHODS FOR THE PRODUCTION OF LIBRARIES FOR DIRECTED EVOLUTION
    44.
    发明公开
    METHODS FOR THE PRODUCTION OF LIBRARIES FOR DIRECTED EVOLUTION 审中-公开
    VERFAHREN ZUR HERSTELLUNG VON BIBLIOTHEKENFÜRDIE GERICHTETE EVOLUTION

    公开(公告)号：EP2961866A1

    公开(公告)日：2016-01-06

    申请号：EP14757218.4

    申请日：2014-02-26

    申请人： Axiomx, Inc.

    发明人： WEINER, Michael ,  KISS, Margaret

    IPC分类号： C40B10/00 ,  C40B40/00 ,  C40B40/06 ,  C40B50/06

    CPC分类号： C12N15/1058 ,  C07K16/00 ,  C07K2317/14 ,  C07K2317/51 ,  C07K2317/515 ,  C07K2317/622 ,  C12N15/1024 ,  C12N15/1037 ,  C12N15/1068 ,  C12N15/1093 ,  C12N15/907 ,  C40B30/04 ,  C40B50/06

    摘要： Disclosed herein is an efficient method of generating a library of variants of a sequence of interest, such as may be used in directed evolution, in one embodiment, the method includes an amplification reaction, e.g. error-prone PCR, to generate double-stranded DNA (dsDNA) variants of a sequence of interest, after which one strand of the dsDNA variants may be selectively degraded to produce single-stranded DNA (ssDNA) variants. The ssDNA variants may be hybridized to ssDNA intermediaries, e.g., uracilated circular ssDNA intermediaries, to form heteroduplex DNA, which may be transformed into cells, such as
    E. coli cells, yielding a library of variants. This method eliminates the inefficient sub-cloning steps and the need for costly primer sets required by many prior methods.

    摘要翻译： 本文公开的是在一个实施方案中产生感兴趣序列的变体文库的有效方法，例如可用于定向进化。该方法包括扩增反应，例如扩增反应。 易发生PCR，以产生目的序列的双链DNA（dsDNA）变体，之后可以选择性降解dsDNA变体的一条链以产生单链DNA（ssDNA）变体。 ssDNA变体可以与ssDNA中间体例如尿嘧啶环状ssDNA中间体杂交以形成异源双链DNA，其可以转化到细胞中，例如大肠杆菌细胞，产生变体文库。 该方法消除了低效的亚克隆步骤以及许多现有方法所需的昂贵的引物组的需要。

    COMPOSITIONS AND METHODS FOR HIGH FIDELITY ASSEMBLY OF NUCLEIC ACIDS
    46.
    发明授权
    COMPOSITIONS AND METHODS FOR HIGH FIDELITY ASSEMBLY OF NUCLEIC ACIDS 有权
    组合物和对于核酸高质量的装配方法

    公开(公告)号：EP2748318B1

    公开(公告)日：2015-11-04

    申请号：EP12753894.0

    申请日：2012-08-23

    申请人： Gen9, Inc.

    发明人： JACOBSON, Joseph ,  SCHINDLER, Daniel ,  LAWTON, Scott S.

    IPC分类号： C12N15/10 ,  C12N15/66 ,  C12P19/34 ,  C40B50/06 ,  G06F19/22

    CPC分类号： C12N15/66 ,  C12N15/10 ,  C12N15/1027 ,  C12N15/1031 ,  C12N15/1089 ,  C12P19/34 ,  C12Q2521/301 ,  C12Q2521/501 ,  G06F19/22

    摘要： Aspects of the invention relate to methods, compositions and algorithms for designing and producing a target nucleic acid. The method can include: (1) providing a plurality of blunt-end double-stranded nucleic acid fragments having a restriction enzyme recognition sequence at both ends thereof; (2) producing via enzymatic digestion a plurality of cohesive-end double-stranded nucleic acid fragments each having two different and non-complementary overhangs; (3) ligating the plurality of cohesive-end double-stranded nucleic acid fragments with a ligase; and (4) forming a linear arrangement of the plurality of cohesive-end double-stranded nucleic acid fragments, wherein the unique arrangement comprises the target nucleic acid. In certain embodiments, the plurality of blunt-end double-stranded nucleic acid fragments can be provided by: releasing a plurality of oligonucleotides synthesized on a solid support; and synthesizing complementary strands of the plurality of oligonucleotides using a polymerase based reaction.

    METHODS FOR NUCLEIC ACID ASSEMBLY AND HIGH THROUGHPUT SEQUENCING
    50.
    发明公开
    METHODS FOR NUCLEIC ACID ASSEMBLY AND HIGH THROUGHPUT SEQUENCING 审中-公开
    方法对于核酸TOGETHER加法和测序高通量

    公开(公告)号：EP2864531A1

    公开(公告)日：2015-04-29

    申请号：EP13809969.2

    申请日：2013-06-24

    申请人： Gen9, Inc.

    发明人： HUDSON, Michael, E. ,  KUNG, Li-jun A. ,  SCHINDLER, Daniel ,  ARCHER, Stephen ,  SAAEM, Ishtiaq

    IPC分类号： C40B50/06

    CPC分类号： C12N15/1031 ,  C12N15/1093 ,  C12N15/66 ,  C12Q2521/301 ,  C12Q2521/501

    摘要： Methods and apparatus of some aspects of the invention relate to the synthesis of high fidelity polynucleotides. In particular, aspects of the invention relate to concurrent enzymatic removal of amplification sequences and ligation of processed oligonucleotides into nucleic acid assemblies. According to some embodiments, the invention provides a method for producing a target nucleic acid having a predefined sequence. In some embodiments, the method comprises the step of providing a plurality of oligonucleotides, wherein each oligonucleotides comprises (i) an internal sequence identical to a different portion of a sequence of a target nucleic acid, (ii) a 5′ sequence flanking the 5′ end of the internal sequence and a 3′ flanking sequence flanking the 3′ end of the internal sequence, each of the flanking sequence comprising a primer recognition site for a primer pair and a restriction enzyme recognition site.