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    • 83. 发明申请
    • METHODS AND SYSTEMS FOR RNA OR DNA DETECTION AND SEQUENCING
    • 用于RNA或DNA检测和测序的方法和系统
    • WO2017184845A1
    • 2017-10-26
    • PCT/US2017/028591
    • 2017-04-20
    • THE ARIZONA BOARD OF REGENTS on behalf of THE UNIVERSITY OF ARIZONA
    • WOROBEY, MichaelWATTS, Thomas
    • C12Q1/68C12Q1/70C07H21/04
    • C12Q1/6844C12Q1/70C12Q1/703C12Q2521/107C12Q2525/204C12Q2537/143
    • Disclosed are methods and systems for detecting RNA and sequencing RNA in a wide range of samples such as samples with low concentrations of nucleic acid, samples with degraded nucleic acid, samples that would not otherwise be amenable to conventional sequencing or RNA detection methods, poor quality samples, high quality samples in which rare mutations are sought, formalin-fixed paraffin-embedded samples, blood samples, etc. The methods of the present invention may use paired, large panels of primers to amplify many short fragments that overlap between but not within each panel. Each panel's amplicon set may fill the gaps between those of the opposing panel, thereby providing complete gene or genomic coverage. A preliminary, multiplex amplification step amplifies target nucleic acid for all downstream reactions such as Sanger sequencing, cloning, and Next Generation Sequencing (NGS).
    • 公开了用于在广泛范围的样品中检测RNA和测序RNA的方法和系统,例如具有低浓度核酸的样品,具有降解的核酸的样品,否则将不适合常规 测序或RNA检测方法,质量差的样品,寻求罕见突变的高质量样品,福尔马林固定的石蜡包埋样品,血液样品等。本发明的方法可以使用成对的大型引物组来扩增许多 在每个面板之间但不在每个面板之间重叠的短片段 每个小组的扩增子组可填补相对小组之间的空白,从而提供完整的基因或基因组覆盖。 初步的多重扩增步骤扩增所有下游反应的目标核酸,如Sanger测序,克隆和下一代测序(NGS)。