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    RNA SEQUENCING AND ANALYSIS USING SOLID SUPPORT
    1.
    发明申请
    RNA SEQUENCING AND ANALYSIS USING SOLID SUPPORT 审中-公开
    使用固体支持的RNA测序和分析

    公开(公告)号：US20100035249A1

    公开(公告)日：2010-02-11

    申请号：US12186009

    申请日：2008-08-05

    申请人： Yoshihide Hayashizaki ,  Piero Carninci ,  Mutsumi Katayama ,  Shintaro Katayama ,  Masayoshi Itoh ,  Matthias Harbers

    发明人： Yoshihide Hayashizaki ,  Piero Carninci ,  Mutsumi Katayama ,  Shintaro Katayama ,  Masayoshi Itoh ,  Matthias Harbers

    IPC分类号： C12Q1/68

    CPC分类号： C12Q1/6809 ,  C12Q1/6834 ,  C12Q1/6874 ,  C12Q2535/101 ,  C12Q2565/537 ,  C12Q2525/155

    摘要： The present invention provides methods for the sequencing of all RNA species within an RNA sample, such as the RNA content obtained from a cell, a tissue, a living organism, or from an artificial source. RNA molecules within the samples are labeled in a RNA-specific manner prior to immobilization on a solid support. One label is used to mark the location of the RNA molecule on the solid support, whereas the second label is used to mark selectively the S′ end of full-length mRNA molecules. RNA molecules are sequenced while being bound to the solid support in one or more sequencing reactions, and sequences of individual RNA molecules can be forwarded to computational analysis for assembling sequence information from individual sequencing reads obtained from the same location on the solid support. Not only unsupervised expression profiling on a genome-wide scale, but also the direct analysis of RNA-RNA interactions become possible as revealed by the analysis of the sequencing information obtained along with genomic information.

    摘要翻译： 本发明提供了用于对RNA样品中所有RNA物种进行测序的方法，例如从细胞，组织，活生物体或人造来源获得的RNA含量。 在固定在固体支持物上之前，将样品中的RNA分子以RNA特异性方式标记。 一个标签用于标记RNA分子在固体支持物上的位置，而第二个标记用于选择性标记全长mRNA分子的S'末端。 RNA分子在一个或多个测序反应中与固体支持物结合时进行测序，并且可将单个RNA分子的序列转发到计算分析，以从固体支持物上的相同位置获得的单独测序读数组装序列信息。 不仅在基因组范围内无监督的表达谱，而且通过分析与基因组信息一起获得的测序信息显示，RNA-RNA相互作用的直接分析也是可能的。

    Cloning vectors and method for molecular cloning
    3.
    发明申请
    Cloning vectors and method for molecular cloning 审中-公开
    克隆载体和分子克隆方法

    公开(公告)号：US20050090010A1

    公开(公告)日：2005-04-28

    申请号：US10469508

    申请日：2002-02-25

    申请人： Yoshihide Hayashizaki ,  Piero Carninci

    发明人： Yoshihide Hayashizaki ,  Piero Carninci

    IPC分类号： C12N15/09 ,  C12N7/00 ,  C12N15/65 ,  C12N15/73 ,  C12P19/34 ,  C12N15/74

    CPC分类号： C12N15/73 ,  C12N15/65

    摘要： The invention discloses a family of cloning vectors capable of cloning nucleic acid inserts of interest of long sizes, with low or reduced background and high efficiency of excision and method for preparing these vectors and library thereof. As example, it is disclosed a cloning vector comprising a construction vector segment (CS) and a replaceable segment (RS), wherein the size of CS is: 36.5 kb≦CS

    摘要翻译： 本发明公开了一种克隆载体，其能够克隆具有低或低背景和高效切割的长尺寸感兴趣的核酸插入物及其制备方法及其文库。 例如，公开了包含构建载体片段（CS）和可替换片段（RS）的克隆载体，其中CS的大小为：36.5kb <= CS <38​​kb，优选CS为37.5kb，包含lox重组 用于网关样重组的Cre重组和/或att重组位点的位点，优选还包括选自以下的背景降低系统：ccdB基因，lox序列，lacZ基因和由限制性内切核酸酶识别的不对称位点序列 。

    Method for modifying RNAS and preparing DNAS from RNAS
    4.
    发明申请
    Method for modifying RNAS and preparing DNAS from RNAS 审中-公开
    RNAS修饰方法及RNAS制备DNAS的方法

    公开(公告)号：US20080108804A1

    公开(公告)日：2008-05-08

    申请号：US11591682

    申请日：2006-11-02

    申请人： Yoshihide Hayashizaki ,  Piero Carninci ,  Matthias Harbers

    发明人： Yoshihide Hayashizaki ,  Piero Carninci ,  Matthias Harbers

    IPC分类号： C07H21/04

    CPC分类号： C07H21/04 ,  C12N15/1096 ,  C12Q1/686 ,  C12Q2521/107 ,  C12Q2533/107

    摘要： A method is disclosed for the modification of an end of RNA molecules and the use of such modified RNA molecules in cDNA synthesis for the purpose of cloning, detection, sequencing, and amplification of parts of the RNAs, the entire RNAs, or any cDNAs derived from such modified RNAs. The invention relates further to the amplification and the identification of nucleic acid molecules for the purpose of single molecule detection and/or high-throughput sequencing. In addition, a method is provided for the preparation of pooled samples that contains molecules each of which is marked by an “Identifier Sequence” for its origin. The invention facilitates studies on biological systems and analysis of genes expressed therein.

    摘要翻译： 公开了用于修饰RNA分子末端的方法以及在cDNA合成中使用这些修饰的RNA分子，以克隆，检测，测序和扩增部分RNA，整个RNA或衍生的任何cDNA 来自这些修饰的RNA。 本发明进一步涉及用于单分子检测和/或高通量测序目的的核酸分子的扩增和鉴定。 此外，提供了一种用于制备包含分子的合并样品的方法，每个分子由其起源的“标识序列”标记。 本发明有助于对生物系统的研究和其中表达的基因的分析。

    Method for isolating DNA
    6.
    发明授权
    Method for isolating DNA 失效
    分离DNA的方法

    公开(公告)号：US06969603B2

    公开(公告)日：2005-11-29

    申请号：US09984742

    申请日：2001-10-31

    申请人： Yoshihide Hayashizaki ,  Piero Carninci

    发明人： Yoshihide Hayashizaki ,  Piero Carninci

    IPC分类号： C12N15/09 ,  C07H21/04 ,  C12N15/10 ,  C12N1/08

    CPC分类号： C12N15/1017 ,  C12N15/1006

    摘要： Method for isolating DNA contained in a biological sample. The method includes combining in a solution a DNA-containing biological sample, a salt, a cationic surfactant, and a DNA-binding carrier, the solution having a salt concentration higher than the DNA precipitation inhibition-initiating concentration, to lyse the DNA-containing biological sample and to bind DNA to the DNA-binding carrier while in the solution to form a bound DNA-carrier. The method also includes separating the DNA-bound carrier from other components. The method further includes dissociating the bound DNA from the DNA-binding carrier. The method still further includes recovering dissociated DNA.

    摘要翻译： 用于分离生物样品中所含DNA的方法。 所述方法包括在溶液中结合含DNA的生物样品，盐，阳离子表面活性剂和DNA结合载体，所述溶液的盐浓度高于DNA沉淀抑制引发浓度，以溶解含DNA的 生物样品，并在溶液中将DNA与DNA结合载体结合，形成结合的DNA载体。 该方法还包括将DNA结合的载体与其它组分分离。 该方法还包括从DNA结合载体解离结合的DNA。 该方法还包括回收解离的DNA。

    Method for Increasing Enzymatic Reactivity
    7.
    发明申请
    Method for Increasing Enzymatic Reactivity 审中-公开
    增加酶活性的方法

    公开(公告)号：US20110059869A1

    公开(公告)日：2011-03-10

    申请号：US12919594

    申请日：2009-02-27

    申请人： Miki Kojima ,  Piero Carninci ,  Yoshihide Hayashizaki ,  Matthias Harbers

    发明人： Miki Kojima ,  Piero Carninci ,  Yoshihide Hayashizaki ,  Matthias Harbers

    IPC分类号： C40B50/06 ,  C12N9/00 ,  C12N9/10 ,  C12N9/14 ,  C12N9/88 ,  C12N9/16 ,  C40B50/14

    CPC分类号： C12N9/96 ,  C12Q1/6834 ,  C12Q1/686 ,  C12Q1/6869 ,  C12Q2527/127 ,  C12Q2527/125 ,  C12Q2521/543

    摘要： An object of the invention is to provide a method for increasing enzymatic reactivity to a target substance immobilized on a support; and a method for reducing or suppressing an inhibitory effect of a support on enzymatic reaction. The above object is achieved by a method for increasing enzymatic reactivity to a target substance immobilized on a support by allowing at least one substance selected from the group consisting of saccharides, amino acids, polyhydric alcohols and derivatives thereof to exist; and a method for reducing or suppressing an inhibitory effect of a support immobilized with a target substance on enzymatic reactivity to the target substance by allowing at least one substance selected from the group consisting of saccharides, amino acids, polyhydric alcohols and derivatives thereof to exist.

    摘要翻译： 本发明的目的是提供一种增加对固定在载体上的目标物质的酶反应性的方法; 以及减少或抑制载体对酶促反应的抑制作用的方法。 上述目的通过使至少一种选自由糖，氨基酸，多元醇及其衍生物组成的组中选出的物质存在而增加对固定在载体上的目标物质的酶反应性的方法来实现。 以及通过使至少一种选自由糖，氨基酸，多元醇及其衍生物组成的组的物质存在的方式，减少或抑制固定有靶物质的载体对目标物质的酶反应性的抑制作用。

    Method for isolating DNA
    9.
    发明授权
    Method for isolating DNA 失效
    分离DNA的方法

    公开(公告)号：US06342387B1

    公开(公告)日：2002-01-29

    申请号：US09508119

    申请日：2000-06-12

    申请人： Yoshihide Hayashizaki ,  Piero Carninci

    发明人： Yoshihide Hayashizaki ,  Piero Carninci

    IPC分类号： C07H108

    CPC分类号： C12N15/1017 ,  C12N15/1006

    摘要： A method for isolating DNA contained in a biological sample, including: lysing a DNA-containing biological sample and forming a DNA-bound carrier by placing a lysing solution, including the DNA-containing biological sample, a salt, and a cationic surfactant, and having a salt concentration higher than the DNA precipitation inhibition-initiating concentration, into contact with a DNA-binding carrier to bind DNA to the DNA-binding carrier to form the DNA-bound carrier; separating the DNA-bound carrier from other components; dissociating the bound DNA from the DNA-binding carrier; and recovering dissociated DNA. By the method, DNA purified with no preliminary treatment of a biological sample can be recovered at a high yield.

    摘要翻译： 一种用于分离生物样品中包含的DNA的方法，包括：通过放置含有DNA的生物样品，盐和阳离子表面活性剂的裂解溶液来裂解含DNA的生物样品并形成DNA结合的载体，以及 具有高于DNA沉淀抑制引发浓度的盐浓度与DNA结合载体接触以将DNA结合到DNA结合载体上以形成DNA结合的载体; 将DNA结合的载体与其他成分分离; 从DNA结合载体解离结合的DNA; 并回收解离的DNA。 通过该方法，可以以高收率回收不经过生物样品的预处理纯化的DNA。

    Method for making full-length cDNA libraries
    10.
    发明授权
    Method for making full-length cDNA libraries 失效
    制备全长cDNA文库的方法

    公开(公告)号：US06174669B1

    公开(公告)日：2001-01-16

    申请号：US08752540

    申请日：1996-11-20

    申请人： Yoshihide Hayashizaki ,  Piero Carninci ,  Claudio Schneider

    发明人： Yoshihide Hayashizaki ,  Piero Carninci ,  Claudio Schneider

    IPC分类号： C12Q168

    CPC分类号： C07H21/00 ,  C07B2200/11 ,  C12N15/1093 ,  C12N15/1096 ,  C40B40/00

    摘要： Disclosed is a method for making full-length CDNA libraries, which is for making libraries of cDNAs having a length corresponding to a full-length of mRNAs and comprises the following steps of; binding a tag molecule to a diol structure present in 5′ Cap (7MeGpppN) sites of mRNAs, forming RNA-DNA hybrids by reverse transcription using primers such as oligo dT and the mRNAs connected with the tag molecule as templates, and separating RNA-DNA hybrids carrying a DNA corresponding to a full-length of mRNAs from the RNA-DNA hybrids formed above by using function of the tag molecule. To obtain mRNA connected with a tag molecule, the diol structure present in 5′ Cap site of mRNA is subjected to a ring-open reaction by oxidation with sodium periodate to form a dialdehyde and the dialdehyde is reacted with a tag molecule having a hydrazine terminus. According to the present invention, there are provided a novel method capable of efficiently labeling 5′ Cap site and a method for making full-length cDNA libraries utilizing the labeling method.

    摘要翻译： 公开了一种制备全长CDNA文库的方法，其用于制备具有对应于全长mRNA的长度的cDNA文库，并且包括以下步骤： 将标签分子与存在于mRNA的5'帽（7MeGpppN）位点的二醇结构结合，通过使用诸如oligo dT的引物和与标签分子连接的mRNA作为模板通过逆转录形成RNA-DNA杂交体，并分离RNA-DNA 通过使用标签分子的功能，携带与来自上述RNA-DNA杂交体的全长mRNA相对应的DNA的杂交体。 为了获得与标签分子连接的mRNA，存在于mRNA的5'帽位点的二醇结构通过用高碘酸钠氧化进行开环反应以形成二醛，并且使二醛与具有肼末端的标签分子反应 。 根据本发明，提供了一种能够利用标记方法有效地标记5'帽位点和制备全长cDNA文库的方法的新方法。