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    SEQUENCER ASSOCIATED WITH A METHOD OF IDENTIFYING DROPLETS IN A STACK
    1.
    发明公开
    SEQUENCER ASSOCIATED WITH A METHOD OF IDENTIFYING DROPLETS IN A STACK 审中-公开

    公开(公告)号：EP3403719A1

    公开(公告)日：2018-11-21

    申请号：EP18183230.4

    申请日：2017-07-14

    申请人： Base4 Innovation Ltd

    发明人： FRAYLING, Cameron Alexander ,  ISAAC, Thomas Henry

    IPC分类号： B01J19/00 ,  B01L3/02 ,  G01N35/10 ,  C12Q1/68 ,  C12Q1/6869

    摘要： Disclosed is an apparatus for sequencing a biopolymer such as a nucleic acid or protein characterised by comprising a digestion unit including an analyte-receiving location where the biopolymer is progressively digested into a corresponding stream of its constituent monomers; a dispensing unit including at least one droplet-dispensing nozzle for dispensing the stream of monomers as a corresponding stream of droplets; a substrate through which is provided an array of open-ended tubes each having an inlet and an outlet on opposing faces and whose major axes are disposed parallel to the direction in which the droplets are to be dispensed; a means for stepping the droplet-dispensing nozzle(s) relative to the inlet face of the substrate in at least one direction perpendicular to the major axes of the tubes thereby allowing the droplets to be dispensed into the tubes; a controller for creating a droplet stack in the tubes for releasing the droplets from the stack through the outlets as required; at least one source of electromagnetic radiation adapted to illuminate the outlets in the outlet face of the substrate; a means for synchronising the illumination of the outlets with the emergence of the droplets therefrom, and; at least one photodetector adapted to detect synchronised fluorescence radiation signals arising from the outlets.

    DROPLET-BASED SEQUENCING METHOD
    2.
    发明公开
    DROPLET-BASED SEQUENCING METHOD 有权
    TRÖPFCHENBASIERTESSEQUENZIERUNGSVERFAHREN

    公开(公告)号：EP2946020A1

    公开(公告)日：2015-11-25

    申请号：EP14700782.7

    申请日：2014-01-17

    申请人： Base4 Innovation Ltd

    发明人： NATALE, Alessandra ,  SOARES, Bruno Flavio Nogueira de Sousa ,  FRAYLING, Cameron Alexander ,  AMASIO, Michele ,  ISAAC, Thomas Henry

    IPC分类号： C12Q1/68

    CPC分类号： G01N21/658 ,  C12Q1/6869 ,  G01N2201/06113 ,  G01N2201/12 ,  C12Q2563/159 ,  C12Q2565/628

    摘要： Disclosed is a method for determining the sequence of nucleotide bases in a polynucleotide analyte. It is characterised by analyte characterised by the steps of: a. generating a stream of droplets at least some of which comprise both (1) a single nucleotide base and (2) colloidal metal particles capable of undergoing plasmon resonance and b. irradiating each droplet with electromagnetic radiation to cause (1) the metal particles also contained therein to undergo plasmon resonance and (2) the nucleotide base contained therein to Raman scatter light at one or more wavelengths characteristic of its type. Suitably, the order of the single nucleotides bases in the droplet stream corresponds to the sequence of nucleotide bases in the analyte.

    摘要翻译： 公开了确定多核苷酸分析物中核苷酸碱基序列的方法。 其特征在于分析物的特征在于以下步骤：a。 产生液滴流，其中至少一些液滴包括（1）单核苷酸碱基和（2）能够进行等离子体共振的胶体金属颗粒和b。 用电磁辐射照射每个液滴，使（1）其中还含有的金属颗粒经历等离子体共振，和（2）其中所含的核苷酸碱基以其类型特征的一个或多个波长的拉曼散射光。 适当地，液滴流中单个核苷酸碱基的顺序对应于分析物中核苷酸碱基的序列。

    SEQUENCING METHOD
    6.
    发明公开
    SEQUENCING METHOD 审中-公开
    SEQUENZIERUNGSVERFAHREN

    公开(公告)号：EP2904109A1

    公开(公告)日：2015-08-12

    申请号：EP13774485.0

    申请日：2013-10-04

    申请人： Base4 Innovation Ltd

    发明人： FRAYLING, Cameron Alexander ,  BALMFORTH, Barnaby ,  SOARES, Bruno Flavio Nogueira de Sousa ,  ISAAC, Thomas Henry ,  BREINER, Boris ,  NATALE, Alessandra ,  AMASIO, Michele

    IPC分类号： C12Q1/68

    CPC分类号： C12Q1/6874 ,  C12Q1/6869 ,  C12Q2521/101 ,  C12Q2521/301 ,  C12Q2521/319 ,  C12Q2521/501 ,  C12Q2525/301 ,  C12Q2563/159 ,  C12Q2565/1015 ,  C12Q2565/629

    摘要： Disclosed is a microfluidic device for sequencing a nucleic acid characterised by comprising: €¢ a first zone in which the analyte is contained and progressively digested into a stream of its constituent single nucleotides; €¢ a primary microfluidic pathway connected to the first zone adapted to allow passage of a carrier solvent containing an ordered stream of primary aqueous microdroplets at least some of which contain one of the single nucleotides; €¢ a second zone disposed within the primary microfluidic pathway and comprising a microdroplet injector and/or a coalescence means for sequentially coalescing secondary aqueous microdroplets from a secondary microfluidic pathway into the primary aqueous microdroplets in the ordered stream; €¢ a storage zone connected to the primary microfluidic pathway downstream of the second zone(s) where the primary aqueous microdroplets are stored; €¢ a light source for interrogating the microdroplets at one or more locations downstream of the primary microfluidic pathway and €¢ a photodetector for detecting fluorescence from the primary aqueous droplets at the location(s).

    摘要翻译： 公开了一种用于对多核苷酸分析物进行测序的方法，包括：a。 产生含有单个核苷酸的液滴流，其中液滴流中单个核苷酸的顺序对应于分析物中的核苷酸序列; b。 向每个液滴中引入多种生物探针类型，每种类型包括不可检测状态的不同标记并适于捕获不同的单个核苷酸; c。 导致液滴中含有的单核苷酸与其互补探针结合， 导致标签从探测器中释放出来，该探针已将核苷酸结合在可检测状态。 探针是一种哑铃形探针，包括荧光供体和猝灭标记以及单个核苷酸间隙。 在通过聚合酶和连接酶修补缺口后，限制酶识别位点被限制酶切割，随后进行核酸外切酶消化以释放标记物。

    DROPLET SEQUENCING DEVICE
    8.
    发明公开
    DROPLET SEQUENCING DEVICE 审中-公开
    滴流测序装置

    公开(公告)号：EP3319729A1

    公开(公告)日：2018-05-16

    申请号：EP16738383.5

    申请日：2016-07-06

    申请人： Base4 Innovation Ltd

    发明人： FRAYLING, Cameron, Alexander ,  ISAAC, Thomas Henry

    IPC分类号： B01L3/02 ,  C12Q1/68 ,  B01L3/00

    摘要： An apparatus for sequencing a nucleic acid by printing droplets at least some of which contain single nucleotides derived from the nucleic acid is provided. It is characterised by comprising; €¢ a planar substrate having a face with droplet-receiving locations arranged in at least one track parallel to a first axis defining the substrate; €¢ a printer head comprising a plurality of droplet-dispensing nozzle types juxtaposed immediately above the droplet-receiving locations so that each nozzle type can in turn dispense a droplet into the droplet-receiving locations; €¢ a means for stepping the printer head along the first axis relative to the droplet-receiving locations; €¢ at least one source of incident electromagnetic radiation adapted to illuminate the droplet-receiving locations and €¢ at least one photodetector to detect adapted to detect fluorescence radiation emitted by each droplet-receiving location after illumination.

    METHOD OF IDENTIFYING DROPLETS IN A STACK AND AN ASSOCIATED SEQUENCER
    10.
    发明公开
    METHOD OF IDENTIFYING DROPLETS IN A STACK AND AN ASSOCIATED SEQUENCER 审中-公开
    确定堆栈中的液滴和相关测序仪的方法

    公开(公告)号：EP3269444A1

    公开(公告)日：2018-01-17

    申请号：EP16179518.2

    申请日：2016-07-14

    申请人： Base4 Innovation Ltd

    发明人： FRAYLING, Cameron Alexander ,  ISAAC, Thomas Henry

    IPC分类号： B01J19/00 ,  B01L3/02 ,  G01N35/10 ,  C12Q1/68

    CPC分类号： B01L3/502784 ,  B01J2219/00367 ,  B01J2219/00576 ,  B01J2219/00702 ,  B01L3/0268 ,  B01L2200/0642 ,  B01L2200/0673 ,  B01L2300/0627 ,  B01L2300/0867 ,  C12Q1/6869 ,  C40B60/12 ,  G01N21/6428 ,  G01N21/6452 ,  G01N2021/6439 ,  C12Q2535/107 ,  C12Q2535/122 ,  C12Q2563/159 ,  C12Q2565/629

    摘要： Disclosed is a method of identifying the contents of individual droplets in a droplet stream each droplet containing fluorophores in an initial non-fluorescing state characterised by the steps of introducing the droplets one-by-one into at least one open-ended tube to create a stack of droplets therein; activating the fluorophores within the droplets to cause them to fluoresce; releasing each droplet in the droplet stack in turn from the tube and detecting along the major axis of the tube fluorescence associated with each droplet as it emerges.
    Also disclosed is a method suitable for sequencing a biopolymer characterised by the steps of (1) progressively digesting the biopolymer into an ordered stream of its constituent monomers; (2) converting the stream of monomers into a corresponding stream of monomer-containing aqueous droplets each droplet additionally containing a probe capable of (a) capturing the monomer and (b) thereafter being digested to release an unqueched fluorophore characteristic of the captured monomer; (3) introducing the stream of droplets created in step (2) into an inlet end of at least one open-ended tube to create a stack of droplets therein and (4) releasing each droplet in turn from an outlet end of the tube(s) and detecting fluorophores in each droplet as each droplet emerges. The method may be used in a corresponding apparatus for sequencing a biopolymer such as a nucleic acid or protein

    摘要翻译： 公开了一种识别液滴流中的单个液滴的内容物的方法，每个液滴包含处于初始非荧光状态的荧光团，其特征在于将所述液滴一个接一个地引入到至少一个开口端管中以产生 其中的小滴堆; 激活液滴内的荧光团以使它们发荧光; 从管中依次释放液滴堆中的每个液滴并随着液滴出现沿着管的主轴检测与每个液滴相关的荧光。 还公开了一种适用于对生物聚合物进行测序的方法，其特征在于以下步骤：（1）逐渐消化生物聚合物为其构成单体的有序流; （2）将单体流转化成含有单体的水性液滴的相应流，每个液滴另外含有能够（a）捕获单体的探针，和（b）随后被消化以释放捕获的单体的未突出的荧光团特征; （3）将步骤（2）中产生的液滴流引入至少一个开口端管的入口端以在其中形成一堆液滴，以及（4）从管的出口端依次释放每个液滴 s）并在每个液滴出现时检测每个液滴中的荧光团。 该方法可用于测定生物聚合物如核酸或蛋白质的相应仪器中