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1. 发明申请

WO2013021175A3 DEVICE AND METHOD FOR PHOTON DETECTION 审中-公开
公开(公告)号：WO2013021175A3
公开(公告)日：2013-02-14
申请号：PCT/GB2012/051873
申请日：2012-08-02
申请人： BASE4 INNOVATION LTD , ISAAC, Thomas Henry
发明人： ISAAC, Thomas Henry
IPC分类号： H03K5/08 , H03K5/15 , H03K5/19 , G01T1/24
摘要： A device (10) for determining the number of discrete events represented by an input signal is provided. The input signal may, for example, comprise pulses representing photons arriving at a detector. The device (10) may comprise a plurality, n, of comparator circuits (14) for reading the signal. For each comparator circuit (14) from i=1 to i=n, the i th comparator circuit (14) has a corresponding threshold value which the amplitude of a pulse representing i discrete events will exceed, but which the amplitude of a pulse representing i-1 discrete events will not exceed. Each comparator circuit (14) is arranged to output a first value when the input signal exceeds its threshold value and a second value when the input signal is less than its threshold value. The device (10) includes a counter (16) for counting the number of outputs of the first value that have been output by the plurality of comparator circuits (14).

2. 发明申请

WO2014167324A1 SINGLE NUCLEOTIDE DETECTION METHOD 审中-公开
标题翻译：单核心检测方法
公开(公告)号：WO2014167324A1
公开(公告)日：2014-10-16
申请号：PCT/GB2014/051106
申请日：2014-04-09
申请人： BASE4 INNOVATION LTD , MEDICAL RESEARCH COUNCIL
发明人： FRAYLING, Cameron Alexander , BALMFORTH, Barnaby , SOARES, Bruno Flavio Nogueira de Sousa , ISAAC, Thomas Henry , BREINER, Boris , NATALE, Alessandra , AMASIO, Michele , DEAR, Paul
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6869 , C12Q2521/319 , C12Q2531/113 , C12Q2537/157 , C12Q2565/629 , C12Q2521/101 , C12Q2521/501 , C12Q2525/301 , C12Q2565/1015 , C12Q2565/301
摘要： A method for determining the sequence of nucleotide bases in a polynucleotide analyte is provided. It is characterised by the steps of (1) generating a stream of single nucleotide bases from the analyte; (2) producing captured molecules by reacting each single nucleotide base with a capture system; (3) amplifying at least part of the captured molecule to produce a plurality of amplicons characteristic of the single nucleotide base; (4) labelling the amplicons with a corresponding probe having a characteristic detectable element and (5) detecting a property characteristic of the detectable element.
摘要翻译：提供了确定多核苷酸分析物中核苷酸碱基序列的方法。其特征在于以下步骤：（1）从分析物产生单核苷酸碱基流; （2）通过使每个单核苷酸碱基与捕获系统反应来产生捕获的分子; （3）扩增至少部分所捕获的分子以产生特征为单核苷酸碱基的多个扩增子; （4）用具有特征可检测元素的相应探针标记扩增子，和（5）检测可检测元件的特性。

3. 发明申请

WO2014167323A1 SINGLE NUCLEOTIDE DETECTION METHOD 审中-公开
标题翻译：单核心检测方法
公开(公告)号：WO2014167323A1
公开(公告)日：2014-10-16
申请号：PCT/GB2014/051105
申请日：2014-04-09
申请人： BASE4 INNOVATION LTD , MEDICAL RESEARCH COUNCIL
发明人： FRAYLING, Cameron Alexander , BALMFORTH, Barnaby , SOARES, Bruno Flavio Nogueira de Sousa , ISAAC, Thomas Henry , BREINER, Boris , NATALE, Alessandra , AMASIO, Michele , DEAR, Paul
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6869 , B01L3/502784 , G01N35/08 , C12Q2521/301 , C12Q2521/319 , C12Q2521/525 , C12Q2525/301 , C12Q2537/157 , C12Q2565/629 , C12Q2521/101 , C12Q2521/501 , C12Q2563/159 , C12Q2565/1015 , C12Q2565/301
摘要： A method for determining the sequence of nucleotide bases in a polynucleotide analyte is provided. It is characterised by the steps of (1) generating a stream of single nucleotide bases from the analyte by pyrophosphorolysis; (2) producing captured molecules by reacting each single nucleotide base with a capture system labelled with detectable elements in an undetectable state; (3) releasing the detectable elements from each captured molecule in a detectable state and (4) detecting the detectable elements so released and determining the sequence of nucleotide bases therefrom. The method can be used advantageously in sequencers involving the use of microdroplets.
摘要翻译：提供了确定多核苷酸分析物中核苷酸碱基序列的方法。其特征在于以下步骤：（1）通过焦磷酸分解从分析物产生单核苷酸碱基流; （2）通过使每个单核苷酸碱基与标记有不可检测状态的可检测元件的捕获系统反应来产生捕获的分子; （3）以可检测状态从每个捕获的分子中释放可检测元素，和（4）检测如此释放的可检测元件，并确定其中的核苷酸碱基序列。该方法可以有利地用于涉及使用微滴的定序器。

4. 发明申请

WO2014053854A1 SEQUENCING METHOD 审中-公开
标题翻译：序列方法
公开(公告)号：WO2014053854A1
公开(公告)日：2014-04-10
申请号：PCT/GB2013/052595
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 method for sequencing a polynucleotide analyte comprising: • a. generating a stream of droplets containing a single nucleotide wherein the order of single nucleotides in the droplet stream corresponds to the sequence of nucleotides in the analyte; • b. introducing into each droplet a plurality of biological probe types each type comprising a different label in an undetectable state and being adapted to capture a different single nucleotide; • c. causing the single nucleotide contained in the droplet to bind to its complementary probe and • d. causing the label to be released from the probe that has bound the nucleotide in a detectable state. The probe is a dumbbell shaped probe comprising fluorescent donor and quencher labels and a single nucleotide gap. After gap repair by a polymerase and a ligase, a restriction enzyme recognition site is cleaved by a restriction enzyme, followed by exonuclease digestion to release the labels.
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，导致标签从已探测状态的核苷酸结合的探针中释放出来。探针是包含荧光供体和猝灭标记和单核苷酸间隙的哑铃形探针。通过聚合酶和连接酶进行间隙修复后，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。

5. 发明申请

WO2014053853A1 BIOLOGICAL PROBES AND THE USE THEREOF 审中-公开
标题翻译：生物探针及其用途
公开(公告)号：WO2014053853A1
公开(公告)日：2014-04-10
申请号：PCT/GB2013/052594
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： C12Q1/6876 , C12Q1/6823 , C12Q1/6869 , C12Q2521/301 , C12Q2521/319 , C12Q2525/301 , C12Q2563/107 , C12Q2565/1015 , C12Q2521/101 , C12Q2521/501
摘要： Disclosed is a biological probe characterised in that it comprises a single-stranded nucleotide region the ends of which are attached to two different oligonucleotide regions wherein at least one of the oligonucleotide regions comprises detectable elements having a characteristic detection property and wherein the detectable elements are so arranged on the oligonucleotide region that the detectable property is less detectable than when the same number detectable elements are bound to a corresponding number of single nucleotides. The biological probe is especially useful for capturing single nucleotides or single-stranded nucleotides to create a used probe which can be degraded by means of a restriction enzyme and an exonuclease to generate single nucleotides carrying a detectable element in a form which can be detected. Typically the detectable elements are fluorophores and the corresponding characteristic fluorescence is rendered undetectable in the probe by for example the use of multiple adjacent fluorophores or mixtures of fluorophores and quenchers attached thereto. Preferably the single stranded nucleotide region is comprised of a single nucleotide whose associated nucleotide base is one of the characteristic of the nucleotides bases found in DNA or RNA.
摘要翻译：公开了一种生物探针，其特征在于其包含单链核苷酸区域，其末端连接到两个不同的寡核苷酸区域，其中至少一个寡核苷酸区域包含具有特征检测性质的可检测元件，并且其中可检测元素为排列在寡核苷酸区域上，当相同数量的可检测元素与相应数量的单个核苷酸结合时，可检测性质较不可检测。生物探针对于捕获单个核苷酸或单链核苷酸特别有用，以产生可以通过限制酶和外切核酸酶降解的用过的探针，以产生携带可检测的形式的可检测元件的单个核苷酸。通常，可检测的元素是荧光团，并且通过例如使用多个相邻的荧光团或与之相连的荧光团和猝灭剂的混合物，在探针中使得相应的特征荧光变得不可检测。优选地，单链核苷酸区域由其相关核苷酸碱基是在DNA或RNA中发现的核苷酸碱基的特征之一的单个核苷酸组成。

6. 发明申请

WO2017005789A1 DROPLET SEQUENCING DEVICE 审中-公开
标题翻译： DROPLET顺序设备
公开(公告)号：WO2017005789A1
公开(公告)日：2017-01-12
申请号：PCT/EP2016/065968
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
CPC分类号： B01L3/0268 , B01L3/5027 , B01L2200/025 , B01L2200/143 , B01L2300/0627 , B01L2300/0819 , C12Q1/6869 , G01N35/1011 , G01N35/1074 , G01N2035/1041 , C12Q2563/107 , C12Q2563/159
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 groups juxtaposed above the droplet-receiving locations so that each nozzle group 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; · a nucleotide-generating site upstream of the printer head by means of which an ordered stream of single nucleotides is created from the nucleic acid and delivered to a first droplet-dispensing nozzle group; • 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.
摘要翻译：提供了通过印刷液滴对核酸进行测序的装置，其中至少一些含有源自核酸的单个核苷酸。其特征在于包括： •平面基板，其具有面部，其具有布置在平行于限定所述基板的第一轴线的至少一个轨道中的液滴接收位置; ·打印头，包括并列在液滴接收位置上方的多个液滴分配喷嘴组，使得每个喷嘴组依次将液滴分配到液滴接收位置; •用于相对于液滴接收位置沿着第一轴线步进打印头的装置; ·在打印机头上游的核苷酸产生位点，通过该位点从核酸产生单个核苷酸的有序流并将其递送到第一液滴分配喷嘴组; •至少一个入射电磁辐射源，其适于照亮所述液滴接收位置;以及·至少一个光电检测器，用于检测适于在照射后检测每个液滴接收位置发射的荧光辐射。

7. 发明申请

WO2014111723A1 DROPLET-BASED SEQUENCING METHOD 审中-公开
标题翻译：基于DROPLET的测序方法
公开(公告)号：WO2014111723A1
公开(公告)日：2014-07-24
申请号：PCT/GB2014/050130
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：披露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）单个核苷酸碱基和（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受等离子体共振，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。

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