公开(公告)号：US20130053280A1

公开(公告)日：2013-02-28

申请号：US13697019

申请日：2011-05-09

申请人： Koshin Hamasaki ,  Toshiro Saito

发明人： Koshin Hamasaki ,  Toshiro Saito

IPC分类号： C40B60/12 ,  C40B40/10 ,  C40B50/14 ,  C40B40/06

CPC分类号： C12Q1/6806 ,  C12Q1/6834 ,  G01N21/05 ,  G01N21/554 ,  G01N21/648 ,  G01N33/54346 ,  C12Q2565/518 ,  C12Q2565/507 ,  C12Q2563/155

摘要： Disclosed is a technique for binding microparticles to patterned bonding pads of a metal (e.g., gold) formed on a support. The microparticles each carry a nucleic acid synthetase or DNA probe immobilized thereon for capturing a nucleic acid sample fragment. The technique involves forming, on a support surface, a film having a thickness equivalent to that of the bonding pads; controlling the size of microparticles with respect to the size of bonding pads; and thereby immobilizing microparticles each bearing a single nucleic acid sample fragment to the bonding pads in a one-to-one manner in a grid form. This allows high-density regular alignment and immobilization of many types of nucleic acid fragment samples on a support and enables high-throughput analysis of nucleic acid samples. Typically, immobilization of microparticles at 1-micrometer intervals easily provides a high density of 106 nucleic acid fragments per square millimeter.

公开(公告)号：US09957562B2

公开(公告)日：2018-05-01

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 ,  G01N21/64 ,  C40B40/06

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.

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。

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.

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。