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    METHOD AND APPARATUS FOR NUCLEIC ACID ANALYSIS
    1.
    发明申请
    METHOD AND APPARATUS FOR NUCLEIC ACID ANALYSIS 审中-公开
    核酸分析方法与装置

    公开(公告)号：US20140200162A1

    公开(公告)日：2014-07-17

    申请号：US14233256

    申请日：2012-05-16

    申请人： Toshiro Saito ,  Koshin Hamasaki ,  Satoshi Takahashi ,  Muneo Maeshima ,  Kyoko Imai ,  Kazumichi Imai ,  Ryuji Tao

    发明人： Toshiro Saito ,  Koshin Hamasaki ,  Satoshi Takahashi ,  Muneo Maeshima ,  Kyoko Imai ,  Kazumichi Imai ,  Ryuji Tao

    IPC分类号： C12Q1/68 ,  C12N15/10

    CPC分类号： C12Q1/6834 ,  C12N15/1065 ,  C12Q2525/207 ,  C12Q2537/125 ,  C12Q2537/143 ,  C12Q2563/143 ,  C12Q2563/149

    摘要： A convenient method for nucleic acid analysis is provided, which enables 1000 or more types of nucleic acid to be analyzed collectively with high comprehensiveness and with a dynamic range of at least four digits. In particular, provided is a very effective analytical method especially for untranslated RNAs and microRNAs, of which the types of target nucleic acids is 10000 or lower. Nucleic acids can be analyzed conveniently and rapidly with high comprehensiveness and quantitative performance at single-molecule sensitivity and resolution by following the steps of: preparing a group of target nucleic acid fragments one molecule at a time and hybridizing the nucleic acid molecules, which have known base sequences and have been labeled with the fluorescence substances, with the group of the target nucleic acid fragments to detect the fluorescence substances labeling the hybridized nucleic acid molecules.

    摘要翻译： 提供了一种方便的核酸分析方法，使得能够以高全面性和至少四位数的动态范围共同分析1000种或更多种类型的核酸。 特别地，提供了非常有效的分析方法，特别是对于非翻译的RNA和微小RNA，其靶核酸的类型为10000或更低。 可以通过以下步骤以单分子灵敏度和分辨率的高全面性和定量性能方便快速地分析核酸：一次一个分子制备一组靶核酸片段，并将已知的核酸分子杂交 碱基序列，并用荧光物质标记，用靶核酸片段组来检测标记杂交核酸分子的荧光物质。

    Method and apparatus for nucleic acid analysis
    2.
    发明授权
    Method and apparatus for nucleic acid analysis 有权

    公开(公告)号：US10294519B2

    公开(公告)日：2019-05-21

    申请号：US14233256

    申请日：2012-05-16

    申请人： Toshiro Saito ,  Koshin Hamasaki ,  Satoshi Takahashi ,  Muneo Maeshima ,  Kyoko Imai ,  Kazumichi Imai ,  Ryuji Tao

    发明人： Toshiro Saito ,  Koshin Hamasaki ,  Satoshi Takahashi ,  Muneo Maeshima ,  Kyoko Imai ,  Kazumichi Imai ,  Ryuji Tao

    IPC分类号： C12Q1/6834 ,  C12N15/10

    摘要： A convenient method for nucleic acid analysis is provided, which enables 1000 or more types of nucleic acid to be analyzed collectively with high comprehensiveness and with a dynamic range of at least four digits. In particular, provided is a very effective analytical method especially for untranslated RNAs and microRNAs, of which the types of target nucleic acids is 10000 or lower. Nucleic acids can be analyzed conveniently and rapidly with high comprehensiveness and quantitative performance at single-molecule sensitivity and resolution by following the steps of: preparing a group of target nucleic acid fragments one molecule at a time and hybridizing the nucleic acid molecules, which have known base sequences and have been labeled with the fluorescence substances, with the group of the target nucleic acid fragments to detect the fluorescence substances labeling the hybridized nucleic acid molecules.

    NUCLEIC ACID ANALYSIS DEVICE, NUCLEIC ACID ANALYSIS APPARATUS, AND NUCLEIC ACID ANALYSIS METHOD
    3.
    发明申请
    NUCLEIC ACID ANALYSIS DEVICE, NUCLEIC ACID ANALYSIS APPARATUS, AND NUCLEIC ACID ANALYSIS METHOD 审中-公开
    核酸分析装置，核酸分析装置和核酸分析方法

    公开(公告)号：US20130157264A1

    公开(公告)日：2013-06-20

    申请号：US13818704

    申请日：2011-07-19

    申请人： Takayuki Obara ,  Kazumichi Imai ,  Toshiro Saito ,  Satoshi Takahashi

    发明人： Takayuki Obara ,  Kazumichi Imai ,  Toshiro Saito ,  Satoshi Takahashi

    IPC分类号： G01N21/64

    CPC分类号： G01N21/6486 ,  C12Q1/6837 ,  G01N21/6428 ,  G01N21/6452 ,  G01N21/648 ,  C12Q2563/173

    摘要： In a nucleic acid analysis device which detects a fluorescent dye on a nucleic acid sample immobilized on a surface of a substrate by exciting the fluorescent dye with an evanescent wave, the detection of a fluorescence signal with a high SN ratio is realized even for a long nucleic acid sample.The nucleic acid analysis device according to the invention is a nucleic acid analysis device in which a plurality of regions for immobilizing a nucleic acid sample are provided on a surface of a support base and a single molecule of a nucleic acid sample is immobilized on at least one of the regions, and which performs sequence determination by performing an extension reaction of the immobilized nucleic acid sample, wherein the immobilization of the single molecule of the nucleic acid sample on the support base is performed at two or more points.

    摘要翻译： 在通过用ev逝波激发荧光染料来检测固定在基板表面上的核酸样品上的荧光染料的核酸分析装置中，即使长时间地实现高SN比的荧光信号的检测 核酸样品。 根据本发明的核酸分析装置是核酸分析装置，其中在支持基底的表面上提供用于固定核酸样品的多个区域，并且将至少一个核酸样品的单个分子固定在至少 其中一个区域，并且通过进行固定化的核酸样品的延伸反应进行序列测定，其中将核酸样品的单分子固定在载体基底上进行两点或更多点。

    NUCLEIC ACID ANALYSIS DEVICE AND NUCLEIC ACID ANALYZER
    6.
    发明申请
    NUCLEIC ACID ANALYSIS DEVICE AND NUCLEIC ACID ANALYZER 有权
    核酸分析装置和核酸分析仪

    公开(公告)号：US20130338041A1

    公开(公告)日：2013-12-19

    申请号：US14002125

    申请日：2012-01-24

    申请人： Koshin Hamasaki ,  Toshiro Saito ,  Takayuki Obara

    发明人： Koshin Hamasaki ,  Toshiro Saito ,  Takayuki Obara

    IPC分类号： C12Q1/68

    CPC分类号： C12Q1/6876 ,  C12Q1/6834 ,  C12Q1/6874 ,  C40B40/06 ,  G01N21/6452 ,  G01N21/648 ,  G01N2021/6439 ,  C12Q2521/543 ,  C12Q2535/122 ,  C12Q2563/149 ,  C12Q2565/513 ,  C12Q2523/308

    摘要： In the conventional nucleic acid analysis devices and nucleic acid analyzers, there was no technique available for sequencing a single nucleic acid molecule easily and highly efficiently. The present invention enabled a highly efficient single molecule immobilization of nucleic acid with good reproductivity in a short time at a low price by providing small metallic bonding pads at predetermined positions on a support substrate, firmly fixing a hydrophobic linker on the bonding pads, and bonding on to the linker bulky microparticles onto which a single molecule of a nucleic acid sample fragment is immobilized. According to the present invention, in the nucleic acid analysis device which uses a nucleic acid analyzer, the nucleotide read length can be extended and many types of nucleic acid molecule to be analyzed can be analyzed at one time.

    摘要翻译： 在常规核酸分析装置和核酸分析仪中，没有技术可以容易且高效率地测序单个核酸分子。 本发明通过在支撑衬底上的预定位置处提供小的金属粘合垫，在接合焊盘上牢固地固定疏水性接头，并且在接合焊盘上牢固地固定疏水性接头，从而能够以低价格在短时间内以高效率的单分子固定具有良好繁殖力的核酸 在其上固定有单个分子的核酸样品片段的连接体庞大的微粒上。 根据本发明，在使用核酸分析仪的核酸分析装置中，可以延长核苷酸读取长度，并且可以一次分析多种类型的要分析的核酸分子。

    DEVICE FOR ANALYZING NUCLEIC ACIDS AND APPARATUS FOR ANALYZING NUCLEIC ACIDS
    7.
    发明申请
    DEVICE FOR ANALYZING NUCLEIC ACIDS AND APPARATUS FOR ANALYZING NUCLEIC ACIDS 审中-公开
    用于分析核酸的装置和分析核酸的装置

    公开(公告)号：US20110281320A1

    公开(公告)日：2011-11-17

    申请号：US13147117

    申请日：2010-01-18

    申请人： Toshiro Saito ,  Kazumichi Imai

    发明人： Toshiro Saito ,  Kazumichi Imai

    IPC分类号： C12N11/14 ,  C12M1/34 ,  C07H21/00 ,  C12M1/40 ,  C12N11/00

    CPC分类号： C12Q1/6816 ,  C12Q2565/50 ,  C12Q2563/143

    摘要： An object of the present invention is to regularly align microparticles, on each of which a nucleic acid synthetase or a DNA probe capable of capturing a nucleic acid sample fragment is immobilized, on a support so as to improve throughput of nucleic acid analysis. The present invention relates to a method comprising immobilizing a nucleic acid synthetase, a DNA probe, or the like in advance to a microparticle, forming a pattern of metal pads each having a diameter smaller than the microparticle diameter with gold or the like on a support, and allowing a microparticle to be bound to the pads via a chemical bond. In addition, when the surfaces of microparticles are electrically charged, a pattern of metal pads each having a diameter equivalent to or larger than the microparticle diameter is formed with gold or the like on a support and a microparticle is allowed to be bound to the pads via a chemical bond. According to the present invention, many types of nucleic acid fragment samples can be regularly aligned at a high density and immobilized on a support. This allows high throughput analysis of nucleic acid samples. For example, if microparticles are immobilized at 1-micron pitches, a high density of 106 nucleic acid fragments/emm2 can be readily achieved.

    摘要翻译： 本发明的一个目的是在载体上规则地将微粒（其中每一个核酸合成酶或能够捕获核酸样品片段的DNA探针）定位在载体上，以便提高核酸分析的生产能力。 本发明涉及一种将核酸合成酶，DNA探针等预先固定在微粒上的方法，在支撑体上形成具有小于微粒径的直径的金属垫的图案 ，并且允许微粒通过化学键与焊盘结合。 此外，当微粒表面带电时，在支撑体上形成金等等于或大于微粒直径的金属垫的图案，并且允许微粒结合到垫 通过化学键。 根据本发明，许多类型的核酸片段样品可以高密度地定期排列并固定在载体上。 这允许核酸样品的高通量分析。 例如，如果微粒以1微米间距固定，则可以容易地实现高密度的106个核酸片段/ emm2。