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    PROCESS FOR AMPLIFYING NUCLEIC ACIDS
    2.
    发明申请
    PROCESS FOR AMPLIFYING NUCLEIC ACIDS 审中-公开
    放大核酸的方法

    公开(公告)号：US20110059868A1

    公开(公告)日：2011-03-10

    申请号：US12855433

    申请日：2010-08-12

    申请人： Yasumasa MITANI ,  Akio YAMANE ,  Yuko SHIBATA ,  Yoshihide HAYASHIZAKI

    发明人： Yasumasa MITANI ,  Akio YAMANE ,  Yuko SHIBATA ,  Yoshihide HAYASHIZAKI

    IPC分类号： C40B40/06 ,  C07H21/04

    CPC分类号： C12Q1/6844 ,  C12Q2525/301

    摘要： The present invention relates to a process for synthesizing or amplifying efficiently a nucleic acid comprising a target nucleic acid sequence. In the process according to the present invention, a primer comprising in its 3′-end portion a sequence (Ac′) which hybridizes a sequence (A) in the 3′-end portion of the target nucleic acid sequence, and in the 5′-side of said sequence (Ac′) a sequence (B′) which hybridizes the complementary sequence (Bc) of a sequence (B) positioned in the 5′-side of said sequence (A) on the target nucleic acid sequence, wherein {X−(Y−Y′)}/X is in the range of −1.00 to 1.00, in which X denotes the number of bases in said sequence (Ac′), Y denotes the number of bases in the region flanked by said sequences (A) and (B) in the target nucleic acid sequence, and Y′ denotes the number of bases in an intervening sequence between said sequences (Ac′) and (B′) (Y′ may be zero).

    摘要翻译： 本发明涉及有效地合成或扩增包含靶核酸序列的核酸的方法。 在本发明的方法中，引物在其3'末端部分包含与目标核酸序列的3'末端部分的序列（A）杂交的序列（Ac'）和5 所述序列（Ac'）的序列（B'）与位于靶核酸序列上的所述序列（A）的5'侧的序列（B）的互补序列（Bc）杂交， 其中{X-（Y-Y'）} / X在-1.00至1.00的范围内，其中X表示所述序列中的碱基数（Ac'），Y表示侧翼的区域中的碱基数 所述序列（A）和（B）在靶核酸序列中，Y'表示所述序列（Ac'）和（B'）之间的间插序列中的碱基数（Y'可以为零）。

    RNA SEQUENCING AND ANALYSIS USING SOLID SUPPORT
    3.
    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。

    Compound, nucleic acid, labeling substance, and detection method
    4.
    发明授权
    Compound, nucleic acid, labeling substance, and detection method 有权

    公开(公告)号：US10294261B2

    公开(公告)日：2019-05-21

    申请号：US14424826

    申请日：2013-09-03

    申请人： KABUSHIKI KAISHA DNAFORM

    发明人： Yoshihide Hayashizaki ,  Takahiro Soma ,  Takeshi Hanami ,  Hajime Kanamori ,  Masaru Baba

    IPC分类号： C07H19/06 ,  C07H21/04 ,  C09B23/04 ,  C09B23/06 ,  C07H19/073 ,  C12Q1/6876

    摘要： The present invention provides a compound represented by the following chemical formula (I); a tautomer or stereoisomer of the compound; or a salt of the compound, the tautomer, or the stereoisomer. In the chemical formula (I), R1 and R2 are each a Group 1 element or a protecting group of an amino group and may be identical to or different from each other, or alternatively, R1 and R2 together may form a protecting group of an amino group. R3 is a Group 1 element or a protecting group of a hydroxy group. R4 is a Group 1 element or —PR5R6R7R8 (R5, R6, R7, and R8 are each a Group 1 element, a lone electron pair, a Group 16 element, a Group 17 element, or a protecting group of a phosphorus atom, and may be identical to or different from each other). J is a hydrogen atom or an arbitrary atomic group, A is a hydrogen atom, a hydroxy group, an alkyl group, an aralkyl group, an alkoxy group, an electron-withdrawing group, a silylene group, or a sulfide group, or alternatively, J and A together may form a linker. L is a single bond or a linker (a linking atom or a linking atomic group), the main chain length (the number of main chain atoms) of the linker is arbitrary, L may or may not contain each of C, N, O, S, P, and Si in the main chain, and may or may not contain each of a single bond, a double bond, a triple bond, an amide bond, an ester bond, a disulfide bond, an imino group, an ether bond, a thioether bond, and a thioester bond in the main chain. Z is an atomic group that can form a peptide bond with a labeling compound, or is an atom or atomic group including a label.

    Pretreatment method of biological sample, detection method of RNA, and pretreatment kit
    6.
    发明授权
    Pretreatment method of biological sample, detection method of RNA, and pretreatment kit 有权
    生物样品预处理方法，RNA检测方法及预处理试剂盒

    公开(公告)号：US09518901B2

    公开(公告)日：2016-12-13

    申请号：US13807605

    申请日：2012-06-28

    申请人： Yoshihide Hayashizaki ,  Kengo Usui ,  Saori Goda ,  Kazuhito Nomura ,  Yuki Kawai

    发明人： Yoshihide Hayashizaki ,  Kengo Usui ,  Saori Goda ,  Kazuhito Nomura ,  Yuki Kawai

    IPC分类号： C12Q1/68 ,  C12P19/34 ,  C12P19/00 ,  C12P19/26 ,  C12P19/28 ,  C12P19/30 ,  G01N1/34 ,  C12N15/10

    CPC分类号： G01N1/34 ,  C12N15/1003 ,  C12Q1/6806

    摘要： The present invention is to provide a pretreatment method that allows RNA to be detected promptly and simply. RNA degradation activity due to lactoferrin present in the human rhinal mucosa is inhibited, for example, by adding iron ion and carbonate ion to a biological sample that contains the human rhinal mucosa. With the pretreated biological sample, an RNA virus gene can be amplified by a reverse transcriptase. Iron ion and carbonate ion can also inhibit reverse transcriptase inhibition due to lysozyme C contained in the human rhinal mucosa. Further, it is preferable to remove the envelope of the RNA virus by adding SDS to the biological sample that contains the human rhinal mucosa.

    摘要翻译： 本发明提供一种能够及时且简单地检测RNA的预处理方法。 例如，通过将铁离子和碳酸根离子加入到含有人类鼻粘膜的生物样品中，可抑制由于存在于人类鼻粘膜中的乳铁蛋白引起的RNA降解活性。 通过预处理的生物样品，RNA病毒基因可以通过逆转录酶扩增。 铁离子和碳酸根离子也可以抑制人类鼻粘膜中包含的溶菌酶C的逆转录酶抑制。 此外，优选通过向包含人类鼻粘膜的生物样品中加入SDS来除去RNA病毒的包膜。

    Method for modifying RNAS and preparing DNAS from RNAS
    7.
    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。

    ISOTHERMAL AMPLIFICATION METHOD AND DNA POLYMERASE USED IN THE SAME
    8.
    发明申请
    ISOTHERMAL AMPLIFICATION METHOD AND DNA POLYMERASE USED IN THE SAME 审中-公开
    同位素放大方法和使用其中的DNA聚合酶

    公开(公告)号：US20100221787A1

    公开(公告)日：2010-09-02

    申请号：US12739496

    申请日：2008-10-24

    申请人： Yoshihide Hayashizaki ,  Masayoshi Itoh ,  Alexander Lezhava ,  Yoshimi Benno ,  Yasumasa Mitani ,  Hajime Kanamori

    发明人： Yoshihide Hayashizaki ,  Masayoshi Itoh ,  Alexander Lezhava ,  Yoshimi Benno ,  Yasumasa Mitani ,  Hajime Kanamori

    IPC分类号： C12P19/34 ,  C12N9/12

    CPC分类号： C12Q1/6846 ,  C12N9/1252 ,  C12P19/34 ,  C12Q2525/155 ,  C12Q2521/107 ,  C12Q2527/101 ,  C12Q2531/107

    摘要： A DNA polymerase suitable for specific isothermal amplification methods and an isothermal amplification method using the DNA polymerase are provided. In the presence of a DNA polymerase including a protein described in the following item (a) or (b), an amplification reaction of a target nucleic acid sequence in a nucleic acid sample is carried out isothermally using a first primer shown in the following (X). By using the DNA polymerase, it becomes possible to carry out the amplification reaction using the primer within a shorter time than ever before. (a) a protein having an amino acid sequence represented by SEQ ID NO. 23 (b) a protein having an amino acid sequence represented by SEQ ID NO. 25 (X) a primer that contains, in a 3′ end portion, a sequence (Ac′) that hybridizes to a sequence (A) of a 3′ end portion of the target nucleic acid sequence and also contains, on a 5′ side of the sequence (Ac′), a sequence (B′) that hybridizes to a complementary sequence (Bc) to a sequence (B) present on a 5′ side with respect to the sequence (A) in the target nucleic acid sequence

    摘要翻译： 提供了适用于特定等温扩增方法的DNA聚合酶和使用DNA聚合酶的等温扩增方法。 在包含下述（a）或（b）中所述的蛋白质的DNA聚合酶的存在下，使用下述第一引物等温进行靶核酸序列的扩增反应 X）。 通过使用DNA聚合酶，可以在比以往更短的时间内使用引物进行扩增反应。 （a）具有SEQ ID NO：1所示的氨基酸序列的蛋白质。 23（b）具有SEQ ID NO：1所示的氨基酸序列的蛋白质。 25（X）引物，其在3'末端含有与目标核酸序列的3'端部分的序列（A）杂交的序列（Ac'），并且在5' 序列（Ac'）侧，与靶序列（A）相对于存在于5'侧的序列（B）的互补序列（Bc）杂交的序列（B'）

    Method for amplifying nucleic acids
    10.
    发明申请
    Method for amplifying nucleic acids 审中-公开
    扩增核酸的方法

    公开(公告)号：US20060110745A1

    公开(公告)日：2006-05-25

    申请号：US11123223

    申请日：2005-05-06

    申请人： Yoshihide Hayashizaki ,  Toshizo Hayashi ,  Yasumasa Mitani

    发明人： Yoshihide Hayashizaki ,  Toshizo Hayashi ,  Yasumasa Mitani

    IPC分类号： C12Q1/68 ,  C12P19/34

    CPC分类号： C12P19/34 ,  C12Q1/6844 ,  C12Q2533/101 ,  C12Q2525/301 ,  C12Q2521/501 ,  C12Q2521/101

    摘要： The invention provides a sequence specific method for amplifying nucleic acids. More particularly, the invention provides a method for amplifying nucleic acid sequences which enables such sequences to be detected with high precision, rapidity and high specificity as compared to conventional methods. The present invention also provides a simple method for cloning nucleic acids, particularly, a rapid and simple method for amplifying alternative splicing forms synthesized by an alternative splicing which is performed in a process of preparing a matured mRNA from a DNA.

    摘要翻译： 本发明提供了用于扩增核酸的序列特异性方法。 更具体地，本发明提供了与常规方法相比，用于扩增能够以高精度，快速和高特异性检测这种序列的核酸序列的方法。 本发明还提供了用于克隆核酸的简单方法，特别是用于扩增通过在从DNA中制备成熟mRNA的过程中进行的选择性剪接合成的可变剪接形式的快速和简单的方法。