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    • 8. 发明申请
    • NUCLEIC ACID AMPLIFICATION WITH INTEGRATED MULTIPLEX DETECTION
    • 具有集成多重检测的核酸扩增
    • WO2006028987A2
    • 2006-03-16
    • PCT/US2005031357
    • 2005-09-02
    • BIOARRAY SOLUTIONS LTDSEUL MICHAELKORZHEVA NATALIYAYANG JIACHENGZHANG YI
    • SEUL MICHAELKORZHEVA NATALIYAYANG JIACHENGZHANG YI
    • C12Q1/68
    • C12Q1/6865C12Q2563/149C12Q2563/107C12Q2537/143
    • A method mediated with in-vitro transcription ("IVT") which permits miniaturization of multiplexed DNA and RN analysis, and in which elongation-mediated multiplexed analysis of polymorphisms (eMAP®) is used as the analysis step, is described. Also described is a method mediated with IVT is for selecting a designated strand from T7­-tagged double a stranded DNA: wherein, the selected strand forms the template for RNA synthesis. In one embodiment, double stranded DNA incorporating the T7 (or other) promoter sequence at the 3' end or the 5'end is produced, for example, by amplification of genomic DNA using the Polymerase Chain Reaction (PCR). Also disclosed are nested PCR designs permitting allele analysis in combination with strand selection by IVT. Further, in one embodiment of a homogeneous format for transcription-mediated amplification and multiplexed detection (which may be particularly suited for viral or pathogen detection), encoded microparticles display "looped" capture probe configurations permitting the generation of a signal upon capture of RNA product and real-time assay monitoring.
    • 描述了使用多体DNA和RN分析进行小型化的体外转录(“IVT”)介导的方法,其中使用延长介导的多态性复合分析(eMAP)作为分析步骤。 还描述了用IVT介导的用于从T7标记的双链DNA选择指定链的方法:其中所选择的链形成用于RNA合成的模板。 在一个实施方案中,例如通过使用聚合酶链反应(PCR)扩增基因组DNA来产生在3'末端或5'端掺入T7(或其它)启动子序列的双链DNA。 还公开了允许等位基因分析与IVT的链选择组合的巢式PCR设计。 此外,在用于转录介导的扩增和多重检测(其可能特别适用于病毒或病原体检测)的均匀形式的一个实施方案中,编码的微粒显示“循环”捕获探针构型,其允许在捕获RNA产物时产生信号 和实时测定监测。