METHODS AND COMPOSITIONS FOR PREPARING RNA FROM A FIXED SAMPLE
    52.
    发明申请
    METHODS AND COMPOSITIONS FOR PREPARING RNA FROM A FIXED SAMPLE 审中-公开
    从固定样品制备RNA的方法和组合物

    公开(公告)号:US20160333394A1

    公开(公告)日:2016-11-17

    申请号:US15162373

    申请日:2016-05-23

    Abstract: The present invention provides improved methods and compositions for RNA isolation. In particular embodiments the present invention concerns the use of methods and compositions for the isolation of full-length RNA from fixed tissue samples. The present invention provides methods for digesting and extracting RNA from a fixed tissue sample.

    Abstract translation: 本发明提供用于RNA分离的改进的方法和组合物。 在具体实施方案中,本发明涉及用于从固定组织样品分离全长RNA的方法和组合物的用途。 本发明提供从固定组织样品中消化和提取RNA的方法。

    Identifying affinity-matured human antibodies
    53.
    发明授权
    Identifying affinity-matured human antibodies 有权
    鉴定亲和力成熟的人抗体

    公开(公告)号:US09453217B2

    公开(公告)日:2016-09-27

    申请号:US14939454

    申请日:2015-11-12

    Applicant: XBiotech, Inc.

    Inventor: John Simard

    Abstract: Antigen-specific immunoglobulin V-regions are identified from a library of nucleic acids amplified using polymerase chain reaction using leader sequence-specific forward primers. The use of leader sequence primers allows all V-region sequences to be amplified (including those with extensive 5′ end mutations) without loss of the original 5′ V gene segment sequence. These libraries can be screened for antigen-specific V-regions using eukaryotic cells engineered to express the amplified V-region-encoding nucleic acids or using bacterial phage display techniques. In the latter, a second V-region library is made using a larger than conventional set of 5′ V-region primers. The sequence errors introduced into the amplification products by this method are corrected using sequence information obtained in the products amplified by the V-region primers to screen the library created using the leader sequence primers. Amino acid sequence information from fragments of donor immunoglobulins can be used to assist in the identification of nucleic acids encoding the heavy and light chains of donor antibodies as well as to design primers to amplify such nucleic acids.

    Abstract translation: 从使用前导序列特异性前向引物的聚合酶链反应扩增的核酸文库鉴定抗原特异性免疫球蛋白V-区。 使用前导序列引物可以扩增所有的V区序列(包括具有5'末端突变的突变),而不损失原始的5'V基因片段序列。 可以使用经工程改造以表达扩增的V区编码核酸的真核细胞或使用细菌噬菌体展示技术来筛选抗原特异性V区的这些文库。 在后者中,使用大于5'V区引物的常规集合制备第二V区文库。 通过该方法引入扩增产物的序列差异使用在由V区引物扩增的产物中获得的序列信息进行校正,以筛选使用前导序列引物产生的文库。 供体免疫球蛋白片段的氨基酸序列信息可用于帮助鉴定编码供体抗体重链和轻链的核酸,以及设计引物以扩增此类核酸。

    METHOD FOR CAPTURING AND ENCODING NUCLEIC ACID FROM A PLURALITY OF SINGLE CELLS
    55.
    发明申请
    METHOD FOR CAPTURING AND ENCODING NUCLEIC ACID FROM A PLURALITY OF SINGLE CELLS 审中-公开
    从多种单细胞中捕获和编码核酸的方法

    公开(公告)号:US20160244742A1

    公开(公告)日:2016-08-25

    申请号:US15025874

    申请日:2014-09-29

    CPC classification number: C12N15/1006 C12N15/1065 C12N15/1096 C12Q2563/185

    Abstract: This invention relates to methods for capturing and encoding nucleic acid from a plurality of single cells. A plurality of solid supports is randomly placed into a plurality of compartments, such that the average number of solid supports per compartment, λ1, is less than 1, wherein each solid support carries (a) a unique identification sequence and (b) a capture moiety. A plurality of single cells is randomly placing into the plurality of compartments, such that the average number of cells per compartment, λ2, is less than 1. These random placement steps may be performed in any order. Nucleic acid is then released from each single cell and captured via the capture moiety, such that nucleic acid from each single cell is tagged with a unique identification sequence.

    Abstract translation: 本发明涉及用于从多个单细胞捕获和编码核酸的方法。 多个固体支持物被随机地放置在多个隔室中,使得每个室的固体支持物的平均数目λ1小于1,其中每个固体支持物携带(a)唯一的识别序列和(b)捕获 部分。 多个单个单元被随机地放置到多个隔室中,使得每隔室的平均单元数λ2小于1.这些随机放置步骤可以以任何顺序执行。 然后从每个单个细胞释放核酸并通过捕获部分捕获,使得来自每个单细胞的核酸用唯一的鉴定序列标记。

    Two-Dimensional Cell Array Device and Apparatus for Gene Quantification and Sequence Analysis
    60.
    发明申请
    Two-Dimensional Cell Array Device and Apparatus for Gene Quantification and Sequence Analysis 审中-公开
    用于基因定量和序列分析的二维细胞阵列装置和装置

    公开(公告)号:US20160010078A1

    公开(公告)日:2016-01-14

    申请号:US14771339

    申请日:2013-03-12

    Applicant: HITACHI, LTD.

    Abstract: In order to conduct gene expression analysis of a number of genes in a number of cells, it has been necessary to separate cells, extract genes therefrom, amplify nucleic acids, and perform sequence analysis. However, separation of cells imposes damages on the cells, and it requires the use of an expensive system. Gene expression analysis in each cell can be carried out with high accuracy by arranging a pair of structures comprising a cell trapping section and a nucleic acid trapping section in a vertical direction to extract individual genes in relevant cells, synthesizing cDNA in the nucleic acid trapping section, amplifying nucleic acids, and analyzing the sequences using a next-generation sequencer.

    Abstract translation: 为了对许多细胞中的许多基因进行基因表达分析,必须分离细胞,从中提取基因,扩增核酸并进行序列分析。 然而,细胞的分离对细胞造成损害,并且需要使用昂贵的系统。 通过在垂直方向布置包含细胞捕获部分和核酸捕获部分的一对结构以提取相关细胞中的单个基因,可以高精度地进行每个细胞中的基因表达分析,在核酸捕获部分中合成cDNA ,扩增核酸,并使用下一代测序仪分析序列。

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