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    SINGLE NUCLEOTIDE DETECTION METHOD
    2.
    发明申请
    SINGLE NUCLEOTIDE DETECTION METHOD 审中-公开

    公开(公告)号：US20180057872A1

    公开(公告)日：2018-03-01

    申请号：US15808004

    申请日：2017-11-09

    申请人： BASE4 INNOVATION LTD

    发明人： Barnaby BALMFORTH ,  Cameron Alexander FRAYLING

    IPC分类号： C12Q1/68

    CPC分类号： C12Q1/6869 ,  C12Q1/6823 ,  C12Q2521/101 ,  C12Q2521/319 ,  C12Q2521/501 ,  C12Q2525/125 ,  C12Q2525/307 ,  C12Q2533/107 ,  C12Q2537/149 ,  C12Q2537/162 ,  C12Q2563/107 ,  C12Q2563/159 ,  C12Q2565/629

    摘要： A method of sequencing a nucleic acid such as DNA or RNA is provided. It is characterised by the steps of (1) generating a stream of single nucleoside triphosphates by progressive pyrophosphorolysis of the nucleic acid; (2) producing at least one substantially double-stranded oligonucleotide used probe by reacting in the presence of a polymerase and a ligase at least one of the single nucleoside triphosphates with a corresponding probe system comprising (a) a first single-stranded oligonucleotide labelled with characteristic detectable elements in an undetectable state and (b) second and third single-stranded oligonucleotides capable of hybridising to complementary regions on the first oligonucleotide; (3) digesting the used probe with an enzyme having double-stranded exonucleolytic activity to yield the detectable elements in a detectable state and a single-stranded fourth oligonucleotide which is at least in part the sequence complement of the first oligonucleotide; (4) reacting the fourth oligonucleotide with another first oligonucleotide to produce a substantially double-stranded oligonucleotide product corresponding to the used probe; (5) repeating steps (3) and (4) in a cycle and (6) detecting the characteristic detectable elements released in each iteration of step (3). Suitably the detectable elements are fluorophores. The method of the present invention generates a stronger fluorescence signal from a single nucleoside triphosphate than has been described previously. Suitable probe systems are also disclosed.

    Isothermal nucleic acid amplification
    3.
    发明授权
    Isothermal nucleic acid amplification 有权

    公开(公告)号：US09657340B2

    公开(公告)日：2017-05-23

    申请号：US14709793

    申请日：2015-05-12

    申请人： ORION PHARMA (UK) LIMITED

    发明人： Mark Jay Hoser

    IPC分类号： C12Q1/68

    CPC分类号： C12P19/34 ,  C12Q1/6844 ,  C12Q2521/507 ,  C12Q2531/119 ,  C12Q2537/162

    摘要： A kit for performing isothermal amplification of a nucleic acid target molecule, where the amplification relies on an upstream primer, a downstream primer, a strand invasion system and an oligonucleotide, wherein the upstream and downstream primers are not substrates for the strand invasion system during the amplification process and do not amplify the target molecule independently of the strand invasion system, wherein the oligonucleotide is a substrate for the strand invasion system.

    MULTIPLEX AMPLIFICATION AND DETECTION
    7.
    发明申请
    MULTIPLEX AMPLIFICATION AND DETECTION 有权
    多功能放大和检测

    公开(公告)号：US20130267434A1

    公开(公告)日：2013-10-10

    申请号：US13822129

    申请日：2011-10-25

    申请人： Guoliang Fu

    发明人： Guoliang Fu

    IPC分类号： C12Q1/68

    CPC分类号： C12Q1/6851 ,  C12Q1/6858 ,  C12Q1/686 ,  C12Q1/6876 ,  C12Q2527/107 ,  C12Q2537/143 ,  C12Q2537/162 ,  C12Q2565/107

    摘要： The invention relates to the field of multiplex amplification. In particular, the invention relates to methods for assaying a sample for one or more nucleic acid targets in a single reaction based on the distinct melting temperatures or melting profiles of primers and/or probes. The invention also provides probes and kits for use in such methods.

    摘要翻译： 本发明涉及多重放大的领域。 特别地，本发明涉及基于引物和/或探针的不同熔解温度或熔融曲线在单一反应中测定一种或多种核酸靶标的样品的方法。 本发明还提供了用于这些方法的探针和试剂盒。

    Methods and compositions for performing low background multiplex nucleic acid amplification reactions
    10.
    发明授权
    Methods and compositions for performing low background multiplex nucleic acid amplification reactions 有权
    用于进行低背景多重核酸扩增反应的方法和组合物

    公开(公告)号：US08293501B2

    公开(公告)日：2012-10-23

    申请号：US12440716

    申请日：2007-09-11

    申请人： Johan Erik Simon Fredriksson ,  Carl Oscar Fredrik Dahl

    发明人： Johan Erik Simon Fredriksson ,  Carl Oscar Fredrik Dahl

    IPC分类号： C12P19/34

    CPC分类号： C12Q1/686 ,  C12Q2537/143 ,  C12Q2521/501 ,  C12Q2537/162 ,  C12Q2525/307 ,  C12Q2531/125

    摘要： Methods and compositions for performing low background multiplex nucleic acid amplification reactions are provided. Aspects of the invention include contacting a nucleic acid sample with two or more primer pairs for two or more target nucleic acids under template dependent primer extension reaction conditions, e.g., polymerase chain reaction (PCR) conditions. The resultant amplified composition is then contacted with target nucleic acid circularizing reagents, and product circularized target nucleic acids are then selected, e.g., for further amplification. Also provided are systems and kits that find use in practicing embodiments of the inventions.

    摘要翻译： 提供了用于进行低背景多重核酸扩增反应的方法和组合物。 本发明的方面包括在模板依赖性引物延伸反应条件例如聚合酶链式反应（PCR）条件下使核酸样品与两种或更多种靶核酸的两个或多个引物对接触。 然后将所得扩增组合物与靶核酸环化试剂接触，然后选择产物环化靶核酸，例如进一步扩增。 还提供了在实践本发明的实施方案中使用的系统和试剂盒。