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    Array-based enzymatic oligonucleotide synthesis
    2.
    发明授权
    Array-based enzymatic oligonucleotide synthesis 有权

    公开(公告)号：US11795450B2

    公开(公告)日：2023-10-24

    申请号：US16563797

    申请日：2019-09-06

    申请人： MICROSOFT TECHNOLOGY LICENSING, LLC

    发明人： Bichlien Nguyen ,  Sergey Yekhanin ,  Karin Strauss

    IPC分类号： C12N15/10 ,  G16B30/00 ,  G16B50/00 ,  C12Q1/6806

    CPC分类号： C12N15/1068 ,  C12Q1/6806 ,  G16B30/00 ,  G16B50/00 ,  C12Q1/6806 ,  C12Q2521/131 ,  C12Q2525/186 ,  C12Q2537/143 ,  C12Q2537/149 ,  C12Q2565/519

    摘要： Array-based enzymatic oligonucleotide synthesis creates a large number of polynucleotides using an uncontrolled and template independent polymerase such as terminal deoxynucleotidyl transferase (TdT). Spatial control of reaction conditions on the surface of the array allows creation of polynucleotides with a variety of arbitrary sequences. Spatial control may be implemented by removing protecting groups attached to nucleotides only at a selected location on the array or by other techniques such as location-specific regulation of enzymatic activity. The ratio of polynucleotides with protecting groups to unprotected polynucleotides used during a cycle of synthesis is adjusted to control the length of homopolymers created by the polymerase. Digital information may be encoded in the enzymatically synthesized polynucleotides. An encoding scheme for representing digital information in a nucleotide sequence accounts for homopolymers in the polynucleotides by collapsing homopolymer strings in the sequence data to a single nucleotide or to a shorter homopolymer.

    SINGLE NUCLEOTIDE DETECTION METHOD
    5.
    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.

    Method for Gene Amplification
    8.
    发明申请
    Method for Gene Amplification 审中-公开
    基因扩增方法

    公开(公告)号：US20170037428A1

    公开(公告)日：2017-02-09

    申请号：US15261274

    申请日：2016-09-09

    申请人： Genodive Pharma Inc.

    发明人： Takashi Horiuchi ,  Takaaki Watanabe

    IPC分类号： C12N15/85

    CPC分类号： C12N15/85 ,  C12N15/63 ,  C12N15/64 ,  C12N2800/30 ,  C12P19/34 ,  C12Q1/6844 ,  C12Q1/6867 ,  C12Q2537/1376 ,  C12Q2525/307 ,  C12Q2537/149 ,  C12Q2521/507

    摘要： The present invention provides a double-stranded DNA constructed specifically for high speed gene amplification, a method for gene amplification and a method for synthesizing protein. The gene amplification system of the present invention used a site-specific recombinase such as Cre-lox system and target sequence thereof to efficiently induce a type of replication referred to as a double rolling-circle replication (DRCR). Amplification unit, whose structure is shown in FIG. 2 (a), is constructed in animal and other cells. DRCR is induced by two recombination events triggered by a site-specific recombinase (Cre) when each replication folk progresses between each pair of target sequences (lox sequences).

    摘要翻译： 本发明提供专门用于高速基因扩增的双链DNA，基因扩增方法和合成蛋白质的方法。 本发明的基因扩增系统使用位点特异性重组酶如Cre-lox系统及其靶序列来有效地诱导称为双滚环复制（DRCR）的复制类型。 放大单元，其结构如图1所示。 2（a），在动物和其他细胞中构建。 当每个复制民族在每对靶序列（lox序列）之间进行时，由位点特异性重组酶（Cre）触发的两个重组事件诱导DRCR。

    Localised RCA-based Amplification Method
    9.
    发明申请
    Localised RCA-based Amplification Method 审中-公开
    基于RCA的局部放大方法

    公开(公告)号：US20160376642A1

    公开(公告)日：2016-12-29

    申请号：US14442701

    申请日：2013-11-14

    申请人： OLINK AB

    发明人： Ulf LANDEGREN ,  Lei CHEN ,  Di WU ,  Yuan NONG ,  Caroline GALLANT

    IPC分类号： C12Q1/68

    CPC分类号： C12Q1/6848 ,  C12Q1/6804 ,  C12Q1/682 ,  C12Q2531/125 ,  C12Q2537/149 ,  C12Q2533/107 ,  C12Q2525/131 ,  C12Q2525/301 ,  C12Q2525/155

    摘要： The present invention provides a method for performing a localised RCA reaction comprising at least two rounds of RCA, wherein the product of a second RCA reaction is attached, and hence localised, to a product of a first RCA reaction, said method comprising: (a) providing a first RCA product; (b) directly or indirectly hybridising to said first RCA product a probe which comprises or provides a primer for a second RCA reaction; and (c) performing a second RCA reaction using said RCA primer of (b) to form a second RCA product, wherein in said reaction: (i) said probe and said primer are not able to prime extension using said first RCA product as template or any such extension is limited to avoid displacement of any probe hybridised to the first RCA product; (ii) the direct or indirect hybridisation of the RCA primer of (b) to the first RCA product is maintained and, by virtue of said hybridisation, the second RCA product is attached to the first RCA product; (iii) a RCA template for said second RCA reaction is comprised in or provided by the probe, or is separately provided. The method finds particular utility in the detection of analytes, wherein the analyte is a nucleic acid or wherein a nucleic acid is used or generated as a marker for the analyte.

    摘要翻译： 本发明提供了一种执行包含至少两轮RCA的局部RCA反应的方法，其中将第二RCA反应的产物连接并因此定位于第一RCA反应的产物，所述方法包括：（a ）提供第一个RCA产品; （b）与所述第一RCA产物直接或间接杂交包含或提供用于第二RCA反应的引物的探针; （c）使用（b）的所述RCA引物进行第二RCA反应以形成第二RCA产物，其中在所述反应中：（i）所述探针和所述引物不能使用所述第一RCA产物作为模板进行扩展 或者任何这样的扩展被限制以避免杂交到第一RCA产品的任何探针的位移; （ii）维持（b）的RCA引物与第一RCA产物的直接或间接杂交，并且由于所述杂交，第二RCA产物连接到第一RCA产物; （iii）用于所述第二RCA反应的RCA模板包含在探针中或由探针提供，或单独提供。 该方法在检测分析物中具有特别的用途，其中分析物是核酸，或者其中使用或产生核酸作为分析物的标记物。

    METHOD FOR DETECTING METHICILLIN-RESISTANT STAPHYLOCOCCUS AUREUS (MRSA) STRAINS
    10.
    发明申请
    METHOD FOR DETECTING METHICILLIN-RESISTANT STAPHYLOCOCCUS AUREUS (MRSA) STRAINS 审中-公开
    检测耐甲氧西林金黄色葡萄球菌（MRSA）菌株的方法

    公开(公告)号：US20150240294A1

    公开(公告)日：2015-08-27

    申请号：US14657148

    申请日：2015-03-13

    申请人： QIAGEN Hamburg GmbH

    发明人： Arnim Wiezer

    IPC分类号： C12Q1/68

    CPC分类号： C12Q1/689 ,  C12Q1/6827 ,  C12Q2537/149

    摘要： The present invention relates, inter alia, to a method for detecting methicillin-resistant Staphylococcus aureus (MRSA) strains, wherein one of the following method variants is carried out: variant A a) chromosomal DNA of S. aureus is isolated from the sample by means of a genome probe, and b) a nucleotide sequence which is specific for MRSA is detected in the isolated DNA; or variant B a) DNA is isolated from the sample, wherein the isolation makes use of a genome probe which is specific for an MRSA nucleotide sequence, preferably an MRSA resistance gene, and b) the DNA isolated in step a) is tested for specific sequences of S. aureus. In addition, suitable kits for carrying out the corresponding methods are provided.

    摘要翻译： 本发明特别涉及检测耐甲氧西林金黄色葡萄球菌（MRSA）菌株的方法，其中进行以下方法变体之一：变体A a）从样品中分离出金黄色葡萄球菌的染色体DNA， 基因组探针的手段，和b）在分离的DNA中检测到对MRSA具有特异性的核苷酸序列; 或变体B a）从样品中分离DNA，其中分离使用对MRSA核苷酸序列特异性的基因组探针，优选MRSA抗性基因，和b）在步骤a）中分离的DNA进行特异性测试 金黄色葡萄球菌的序列。 另外，提供了用于实施相应方法的合适的试剂盒。