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    Polymerase idling method for single molecule DNA sequencing
    11.
    发明授权
    Polymerase idling method for single molecule DNA sequencing 有权
    用于单分子DNA测序的聚合酶空转方法

    公开(公告)号：US09546398B2

    公开(公告)日：2017-01-17

    申请号：US14538632

    申请日：2014-11-11

    Applicant: Agilent Technologies, Inc.

    Inventor： Brian Jon Peter ,  John Mannion ,  Joel Myerson

    IPC: C12Q1/68

    CPC classification number: C12Q1/6869 ,  C12Q1/6816 ,  C12Q1/6834 ,  C12Q2525/125 ,  C12Q2525/186 ,  C12Q2537/157

    Abstract: A method for sequencing a nucleic acid is provided. In certain embodiments the method comprises obtaining a duplex comprising a nucleic acid and a primer, wherein the primer has a nuclease resistant 3′ end, combining the duplex with a chain terminator nucleotide and a proof-reading polymerase to produce a reaction in which the polymerase idles on the added chain terminator nucleotide, identifying the chain terminator nucleotide added to the end of the primer; and adding a nuclease-resistant nucleotide to the end of the primer after the polymerase has idled on and removed the added chain terminator nucleotide, thereby producing a duplex comprising the template and an extended primer that has a nuclease resistant 3′ end.

    Abstract translation: 提供了一种核酸测序方法。 在某些实施方案中，该方法包括获得包含核酸和引物的双链体，其中引物具有核酸酶抗性3'末端，将双链体与链终止子核苷酸和校对阅读聚合酶组合以产生其中聚合酶 在添加的链终止子核苷酸上空转，鉴定添加到引物末端的链终止子核苷酸; 并在聚合酶空转之后向引物的末端添加核酸酶抗性核苷酸并除去加入的链终止子核苷酸，从而产生包含模板的双链体和具有核酸酶抗性3'末端的延伸引物。

    POLYMERASE IDLING METHOD FOR SINGLE MOLECULE DNA SEQUENCING
    12.
    发明申请
    POLYMERASE IDLING METHOD FOR SINGLE MOLECULE DNA SEQUENCING 有权
    用于单分子DNA测序的聚合酶识别方法

    公开(公告)号：US20150132756A1

    公开(公告)日：2015-05-14

    申请号：US14538632

    申请日：2014-11-11

    Applicant: Agilent Technologies, Inc.

    Inventor： Brian Jon Peter ,  John Mannion ,  Joel Myerson

    IPC: C12Q1/68

    CPC classification number: C12Q1/6869 ,  C12Q1/6816 ,  C12Q1/6834 ,  C12Q2525/125 ,  C12Q2525/186 ,  C12Q2537/157

    Abstract: A method for sequencing a nucleic acid is provided. In certain embodiments the method comprises obtaining a duplex comprising a nucleic acid and a primer, wherein the primer has a nuclease resistant 3′ end, combining the duplex with a chain terminator nucleotide and a proof-reading polymerase to produce a reaction in which the polymerase idles on the added chain terminator nucleotide, identifying the chain terminator nucleotide added to the end of the primer; and adding a nuclease-resistant nucleotide to the end of the primer after the polymerase has idled on and removed the added chain terminator nucleotide, thereby producing a duplex comprising the template and an extended primer that has a nuclease resistant 3′ end.

    Abstract translation: 提供了对核酸进行测序的方法。 在某些实施方案中，该方法包括获得包含核酸和引物的双链体，其中引物具有核酸酶抗性3'末端，将双链体与链终止子核苷酸和校对阅读聚合酶组合以产生其中聚合酶 在添加的链终止子核苷酸上空转，鉴定添加到引物末端的链终止子核苷酸; 并在聚合酶空转之后向引物的末端添加核酸酶抗性核苷酸并除去加入的链终止子核苷酸，从而产生包含模板的双链体和具有核酸酶抗性3'末端的延伸引物。

    C-probe libraries for DNA target enrichment
    15.
    发明授权
    C-probe libraries for DNA target enrichment 有权

    公开(公告)号：US10150960B2

    公开(公告)日：2018-12-11

    申请号：US13777842

    申请日：2013-02-26

    Applicant: Agilent Technologies, Inc.

    Inventor： Brian Jon Peter

    IPC: C12Q1/68 ,  C12N9/00 ,  C07H21/04 ,  C12N15/10

    Abstract: Provided herein is a method for enriching a target nucleic acid molecule. In one embodiment, the method may involve hybridizing a C-probe to a strand of a target nucleic acid to produce a complex, enzymatically removing any 3′ overhanging end from the target nucleic acid of the complex to produce a 3′ hydroxyl group at the 3′ end; extending the 3′ end of the first sequence using the oligonucleotide sequence of the C-probe as a template; enzymatically removing any 5′ overhanging end from the target nucleic acid, either before or after the extending step, to produce an 5′ phosphate group at the end of the second sequence; and ligating the 5′ phosphate group at the end of the second sequence to the 3′ hydroxyl group at the end of the first sequence to produce a circular DNA molecule that contains the target sequence and the complement of the oligonucleotide sequence.

    HAIRPIN-MEDIATED AMPLIFICATION METHOD
    16.
    发明申请
    HAIRPIN-MEDIATED AMPLIFICATION METHOD 审中-公开

    公开(公告)号：US20180073068A1

    公开(公告)日：2018-03-15

    申请号：US15692215

    申请日：2017-08-31

    Applicant: Agilent Technologies, Inc.

    Inventor： Brian Jon Peter ,  Robert A. Ach

    IPC: C12Q1/68

    CPC classification number: C12Q1/6853 ,  C12Q1/6806 ,  C12Q2521/501 ,  C12Q2525/151 ,  C12Q2525/301

    Abstract: In some embodiments, the amplification method may comprise producing a reaction mix comprising: a nucleic acid sample, a polymerase, nucleotides, a forward primer that hybridizes to a sequence in the bottom strand of a fragment in the sample, and a reverse primer. The reverse primer has a hairpin structure comprising a loop, a stem and a 3′ overhang of at least 8 nucleotides, wherein the 3′ overhang hybridizes to a sequence in the top strand of the fragment. Subjecting the reaction mix at least two rounds of denaturation, renaturation and primer extension conditions results in extension the forward and reverse primers to produce an amplification product that contains: a double stranded region comprising a nick adjacent to the 5′ end of the reverse primer, and the loop of the first hairpin primer. Primer sets and kits for performing the methods are also provided.

    SPATIAL MOLECULAR BARCODING OF IN SITU NUCLEIC ACIDS
    18.
    发明申请
    SPATIAL MOLECULAR BARCODING OF IN SITU NUCLEIC ACIDS 有权
    原子核酸的空间分子束缚

    公开(公告)号：US20150148239A1

    公开(公告)日：2015-05-28

    申请号：US14493127

    申请日：2014-09-22

    Applicant: Agilent Technologies, Inc.

    Inventor： Brian Jon Peter ,  Robert A. Ach ,  Alicia Scheffer-Wong ,  Carolina Caffaro

    IPC: C12Q1/68

    CPC classification number: C12Q1/6841 ,  C12Q2565/513 ,  C12Q2565/514

    Abstract: This disclosure provides, among other things, a method for analyzing a planar cellular sample. In some embodiments, the method comprises: (a) indirectly or directly attaching nucleic acid tags to binding sites in a planar cellular sample; (b) contacting the planar cellular sample with a solid support comprising an array of spatially addressed features that comprise oligonucleotides, wherein each oligonucleotide comprises a molecular barcode that identifies the feature in which the oligonucleotides is present; (c) hybridizing the nucleic acid tags, or a copy of the same, with the oligonucleotides to produce duplexes; and (d) extending the oligonucleotides in the duplexes to produce extension products that each comprises (i) a molecular barcode and (ii) a copy of a nucleic acid tag. Other embodiments, e.g., kits and the like, are also described.

    Abstract translation: 本公开尤其提供了一种用于分析平面细胞样品的方法。 在一些实施方案中，所述方法包括：（a）将核酸标签间接或直接附着于平面细胞样品中的结合位点; （b）使所述平面细胞样品与包含寡核苷酸的空间寻址特征阵列的固体支持物接触，其中每个寡核苷酸包含鉴定存在寡核苷酸的特征的分子条形码; （c）将核酸标签或其拷贝与寡核苷酸杂交以产生双链体; 和（d）在双链体中延伸寡核苷酸以产生延伸产物，其各自包含（i）分子条形码和（ii）核酸标签的拷贝。 还描述了其它实施例，例如套件等。

    Nanofluidic Device for Charge Analysis of Straightened Molecules
    19.
    发明申请
    Nanofluidic Device for Charge Analysis of Straightened Molecules 有权
    用于直线分子电荷分析的纳米流体装置

    公开(公告)号：US20150068901A1

    公开(公告)日：2015-03-12

    申请号：US14482803

    申请日：2014-09-10

    Applicant: Agilent Technologies, Inc.

    Inventor： John Mannion ,  Bo Curry ,  Brian Jon Peter

    IPC: G01N27/447

    CPC classification number: G01N27/4145 ,  G01N27/4473

    Abstract: This disclosure provides, among other things, a nanofluidic device sensing device is provided. In certain embodiments, the device contains: a) a channel comprising a floor and a ceiling, b) an array of charge sensors in the floor and/or ceiling of the channel, arranged along the longitudinal axis of the channel; c) a capture area in the floor and/or ceiling of the channel at the entrance end of the channel; and d) a first electrode and a second electrode, wherein the first and second electrodes are positioned to provide an electrophoretic force along the longitudinal axis of the channel. Other embodiments, e.g., methods, are also described.

    Abstract translation: 本公开尤其提供了一种纳米流体装置感测装置。 在某些实施例中，该装置包括：a）包括地板和天花板的通道，b）沿通道的纵向轴线布置的通道的地板和/或天花板中的一组电荷传感器; c）在通道的入口端的通道的地板和/或天花板中的捕获区域; 以及d）第一电极和第二电极，其中所述第一和第二电极被定位成沿着所述通道的纵向轴线提供电泳力。 还描述了其它实施例，例如方法。

    NUCLEIC ACID ENRICHMENT USING CAS9
    20.
    发明申请
    NUCLEIC ACID ENRICHMENT USING CAS9 审中-公开
    使用CAS9的核酸增殖

    公开(公告)号：US20140356867A1

    公开(公告)日：2014-12-04

    申请号：US14290901

    申请日：2014-05-29

    Applicant: Agilent Technologies, Inc.

    Inventor： Brian Jon Peter ,  Robert A. Ach

    IPC: C12Q1/68 ,  C12N15/10

    Abstract: A method of enriching for a fragment of a genome, as well as corresponding compositions and kits, are provided. In certain embodiments, the method comprises: (a) contacting a sample comprising fragmented DNA with a Cas9-gRNA complex comprising mutant Cas9 protein that has inactivated nuclease activity and a Cas9-associated guide RNA that is complementary to a site in the DNA, to produce a Cas9-fragment complex that comprises a fragment of the fragmented DNA; and (b) isolating the complex. In addition, other methods and compositions for Cas9/CRISPR-mediated nucleic acid manipulation are also provided.

    Abstract translation: 提供了富集基因组片段的方法，以及相应的组合物和试剂盒。 在某些实施方案中，所述方法包括：（a）使包含片段化DNA的样品与Cas9-gRNA复合物接触，所述Cas9-gRNA复合物包含具有失活的核酸酶活性的突变型Cas9蛋白和与DNA中的位点互补的Cas9相关导向RNA， 产生包含片段化DNA的片段的Cas9片段复合体; 和（b）分离复合物。 此外，还提供了用于Cas9 / CRISPR介导的核酸操作的其它方法和组合物。

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