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    • 3. 发明申请
    • METHOD AND PRODUCT FOR LOCALIZED OR SPATIAL DETECTION OF NUCLEIC ACID IN A TISSUE SAMPLE
    • 在组织样品中局部或空间检测核酸的方法和产品
    • US20140066318A1
    • 2014-03-06
    • US14111482
    • 2012-03-13
    • Jonas FrisenPatrik StåhlJoakim Lundeberg
    • Jonas FrisenPatrik StåhlJoakim Lundeberg
    • C12Q1/68
    • The present invention relates to methods and products for the localized or spatial detection of nucleic acid in a tissue sample and in particular to a method for localized detection of nucleic acid in a tissue sample comprising: (a) providing an array comprising a substrate on which multiple species of capture probes are directly or indirectly immobilized such that each species occupies a distinct position on the array and is oriented to have a free 3′ end to enable said probe to function as a primer for a primer extension or ligation reaction, wherein each species of said capture probe comprises a nucleic acid molecule with 5′ to 3′: (i) a positional domain that corresponds to the position of the capture probe on the array, and (ii) a capture domain; (b) contacting said array with a tissue sample such that the position of a capture probe on the array may be correlated with a position in the tissue sample and allowing nucleic acid of the tissue sample to hybridize to the capture domain in said capture probes; (c) generating DNA molecules from the captured nucleic acid molecules using said capture probes as extension or ligation primers, wherein said extended or ligated DNA molecules are tagged by virtue of the positional domain; (d) optionally generating a complementary strand of said tagged DNA and/or optionally amplifying said tagged DNA; (e) releasing at least part of the tagged DNA molecules and/or their complements or amplicons from the surface of the array, wherein said part includes the positional domain or a complement thereof; and (f) directly or indirectly analyzing the sequence of the released DNA molecules.
    • 本发明涉及用于组织样本中核酸的局部或空间检测的方法和产品,特别涉及组织样品中核酸的局部检测方法,包括:(a)提供包含基质的阵列, 直接或间接固定多种捕获探针,使得每个物种在阵列上占据不同的位置并且被定向为具有游离的3'末端以使得所述探针能够用作引物延伸或连接反应的引物,其中每个 所述捕获探针的种类包括具有5'至3'的核酸分子:(i)对应于阵列上的捕获探针的位置的位置结构域,和(ii)捕获结构域; (b)使所述阵列与组织样品接触,使得阵列上的捕获探针的位置可与组织样品中的位置相关,并允许组织样品的核酸与所述捕获探针中的捕获结构域杂交; (c)使用所述捕获探针作为延伸或连接引物从捕获的核酸分子产生DNA分子,其中所述延伸或连接的DNA分子由于位置结构域而被标记; (d)任选地产生所述标记的DNA的互补链和/或任选地扩增所述标记的DNA; (e)从阵列的表面释放标记的DNA分子和/或其补体或扩增子的至少一部分,其中所述部分包括位置结构域或其互补体; 和(f)直接或间接分析释放的DNA分子的序列。
    • 5. 发明授权
    • Method and product for localized or spatial detection of nucleic acid in a tissue sample
    • US10030261B2
    • 2018-07-24
    • US14111482
    • 2012-04-13
    • Jonas FrisenPatrik StåhlJoakim Lundeberg
    • Jonas FrisenPatrik StåhlJoakim Lundeberg
    • C12Q1/6837C12Q1/6834C40B20/02C12Q1/6841C12Q1/6844
    • The present invention relates to methods and products for the localized or spatial detection of nucleic acid in a tissue sample and in particular to a method for localized detection of nucleic acid in a tissue sample comprising: (a) providing an array comprising a substrate on which multiple species of capture probes are directly or indirectly immobilized such that each species occupies a distinct position on the array and is oriented to have a free 3′ end to enable said probe to function as a primer for a primer extension or ligation reaction, wherein each species of said capture probe comprises a nucleic acid molecule with 5′ to 3′: (i) a positional domain that corresponds to the position of the capture probe on the array, and (ii) a capture domain; (b) contacting said array with a tissue sample such that the position of a capture probe on the array may be correlated with a position in the tissue sample and allowing nucleic acid of the tissue sample to hybridize to the capture domain in said capture probes; (c) generating DNA molecules from the captured nucleic acid molecules using said capture probes as extension or ligation primers, wherein said extended or ligated DNA molecules are tagged by virtue of the positional domain; (d) optionally generating a complementary strand of said tagged DNA and/or optionally amplifying said tagged DNA; (e) releasing at least part of the tagged DNA molecules and/or their complements or amplicons from the surface of the array, wherein said part includes the positional domain or a complement thereof; and (f) directly or indirectly analyzing the sequence of the released DNA molecules.