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    • 4. 发明申请
    • CONTROLLING FLUORESCENT SIGNAL COMPOSITION IN OPTICALLY-BASED NANOPORE SEQUENCING
    • 控制基于光学的纳米孔测序中的荧光信号组成
    • WO2018071273A1
    • 2018-04-19
    • PCT/US2017/055435
    • 2017-10-05
    • QUANTAPORE, INC.
    • HUBER, Martin
    • C12Q1/68G01N21/64G01N33/487G01N21/76
    • The invention is directed to optically based methods for nanopore sequencing of polynucleotides which comprise steps of (i) translocating a polynucleotide through a bore of a nanopore at a translocation speed, wherein nucleotides of the polynucleotide are labeled with fluorescent labels such that in free solution fluorescent labels of nucleotides are substantially quenched and wherein fluorescent labels within the bore are constrained such that substantially no detectable fluorescent signal is generated therein; (ii) exciting the fluorescent label of each nucleotide upon exiting the nanopore and prior to quenching with a preceding mutually quenching fluorescent label or a quenching agent; (iii) measuring a fluorescent signal generated by fluorescent labels exiting the nanopore, wherein the translocation speed is selected so that the measured fluorescent signal comprises fluorescence from substantially a single fluorescent label; and (iv) determining a nucleotide sequence of the polynucleotide from a sequence of measured fluorescent signals.
    • 本发明涉及用于多核苷酸的纳米孔测序的基于光学的方法,其包括以下步骤:(i)以易位速度使多核苷酸易位穿过纳米孔的孔,其中多核苷酸的核苷酸用 荧光标记,使得在游离溶液中核苷酸的荧光标记基本上被淬灭,并且其中孔内的荧光标记受到限制,使得其中基本上不产生可检测的荧光信号; (ii)激发离开纳米孔后每种核苷酸的荧光标记并在用先前的相互猝灭荧光标记或猝灭剂猝灭之前; (iii)测量由离开纳米孔的荧光标记产生的荧光信号,其中选择易位速度使得测量的荧光信号包括来自基本上单个荧光标记的荧光; 和(iv)从测量的荧光信号序列确定多核苷酸的核苷酸序列。
    • 6. 发明申请
    • OPTICALLY BASED NANOPORE SEQUENCING
    • 基于光学的纳米孔测序
    • WO2018009346A1
    • 2018-01-11
    • PCT/US2017/038813
    • 2017-06-22
    • QUANTAPORE, INC.
    • ANDERSON, Brett N.
    • G01N27/447C12Q1/68
    • In some aspects the invention is directed to methods of analyzing a polynucleotide which include steps of directing to a nanopore an excitation beam having a predetermined polarization state; translocating a polynucleotide through the nanopore, wherein nucleotides of the polynucleotide are labeled with fluorescent labels having absorption dipoles and wherein the nanopore spatially orients the fluorescent labels so that during translocation the adsorption dipoles are substantially unresponsive to the excitation beam; detecting changes in fluorescent signals generated by the fluorescent labels as nucleotides with fluorescent labels exit the nanopore and absorption dipoles thereof become responsive to excitation by the excitation beam with the predetermined polarization state; and identifying nucleotides exiting the nanopore from the changes in fluorescent signals.
    • 在一些方面,本发明涉及分析多核苷酸的方法,所述方法包括以下步骤:将具有预定偏振态的激发束引导至纳米孔; 使多核苷酸转位穿过纳米孔,其中多核苷酸的核苷酸用具有吸收偶极子的荧光标记标记,并且其中纳米孔在空间上定向荧光标记,使得在易位过程中吸附偶极对激发束基本上没有反应; 检测荧光标记产生的荧光信号的变化,如荧光标记的核苷酸离开纳米孔并且其吸收偶极子变得响应于具有预定偏振状态的激发束的激励; 并从荧光信号的变化中鉴定离开纳米孔的核苷酸。
    • 7. 发明申请
    • MIXED OPTICAL SIGNALS IN POLYMER ANALYSIS WITH NANOPORES
    • 纳米粒子在聚合物分析中的混合光学信号
    • WO2017180319A1
    • 2017-10-19
    • PCT/US2017/024314
    • 2017-03-27
    • QUANTAPORE, INC.
    • ANDERSON, Brett N.HUBER, MartinMACEVICZ, Stephen C.
    • C12Q1/68C40B20/00
    • The invention is directed to nanopore-based methods for analyzing polymers, such as polynucleotides or proteins, containing optical labels specific for different kinds of monomers. In some embodiments, methods of the invention include steps of (a) translocating a polymer through a nanopore, wherein different kinds of monomers of the polymer are labeled with different optical labels that generate distinguishable optical signals and wherein the nanopore constrains the monomers to move single file through an excitation zone that encompasses a plurality of monomers; (b) detecting a time-ordered set of optical signals from the monomers as the polymer passes through the excitation zone; (c) separating optical signals from different kinds of monomers to form monomer-specific time-ordered sets of optical signals; and (d) determining a sequence of monomers from the monomer-specific time-ordered sets of optical signals from the polymer.
    • 本发明涉及用于分析聚合物(例如多核苷酸或蛋白质)的基于纳米孔的方法,所述聚合物含有对不同种类的单体特异的光学标记。 在一些实施方案中,本发明的方法包括以下步骤:(a)使聚合物易位穿过纳米孔,其中聚合物的不同种类的单体用产生可区分的光信号的不同光学标记标记,并且其中纳米孔约束单体以移动单个 通过包含多种单体的激发区进行培养; (b)当聚合物通过激发区时,检测来自单体的时间顺序的一组光信号; (c)从不同种类的单体分离光学信号以形成单体特定的时间有序的光学信号组; 和(d)从来自聚合物的单体特异性时间顺序的光学信号组中确定单体的顺序。