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    • 3. 发明申请
    • NUCLEIC ACID DETECTION AND QUANTIFICATION BY POST-HYBRIDIZATION LABELING AND UNIVERSAL ENCODING
    • 通过后期杂交标签和通用编码进行核酸检测和定量
    • WO2011156434A2
    • 2011-12-15
    • PCT/US2011/039531
    • 2011-06-07
    • FIREFLY BIOWORKS, INC.PREGIBON, Daniel, C.STONER, IsaacWINDEMUTH, AndreasERPS, Timothy
    • PREGIBON, Daniel, C.STONER, IsaacWINDEMUTH, AndreasERPS, Timothy
    • C12Q1/68C12N15/11G01N33/53
    • C12Q1/6825C12Q1/6809C12Q1/6813C12Q1/6816C12Q1/6834C12Q1/6888C12Q1/689C12Q1/6895C12Q1/70G01N15/14G01N21/47G01N21/6428G01N33/6803C12Q2565/626C12Q2525/161C12Q2525/191C12Q2533/107
    • The present invention provides, among other things, methods and compositions for detecting and quantifying target nucleic acids via post-hybridization labeling. In some embodiments, the present invention provides a method for detecting the presence and/or abundance of target nucleic acids in a sample by (a) contacting a plurality of nucleic acid probes with a sample, each nucleic acid probe comprising a capturing sequence for binding a target nucleic acid and an adjacent adapter sequence for binding a universal adapter; (b) incubating the plurality of probes and the sample, in the presence of one or more universal adapters, under conditions that permit binding of both an individual target nucleic acid and an individual universal adapter to a same individual nucleic acid probe; (c) carrying out a reaction that allows coupling of the individual universal adapter to the individual target nucleic acid when hybridized to the same individual nucleic acid probe; (d) detecting the presence of the one or more universal adapters associated with the plurality of nucleic acid probes, thereby detecting the presence of the target nucleic acids in the sample. Typically, universal adapters are labeled with detectable moieties or other labeling groups to facilitate detection. In some embodiments, the plurality of nucleic acid probes suitable for the invention are attached to a substrate or object (e.g., microarray, or particle).
    • 本发明尤其提供了用于通过后杂交标记检测和定量靶核酸的方法和组合物。 在一些实施方案中,本发明提供了通过(a)使多个核酸探针与样品接触来检测样品中靶核酸的存在和/或丰度的方法,每个核酸探针包含用于结合的捕获序列 目标核酸和用于结合通用适配器的相邻适配器序列; (b)在一种或多种通用衔接子的存在下,在允许单独的靶核酸和单个通用衔接子与相同的单个核酸探针结合的条件下孵育多个探针和样品; (c)当与相同的单个核酸探针杂交时进行允许单个通用衔接子与单个靶核酸偶联的反应; (d)检测与多个核酸探针相关联的一个或多个通用适配器的存在,从而检测样品中靶核酸的存在。 通常,通用衔接子用可检测部分或其他标记基团标记以便于检测。 在一些实施方案中,适用于本发明的多个核酸探针连接到底物或物体(例如微阵列或颗粒)上。