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    Methods for Amplification of Nucleic Acids Utilizing Clamp Oligonucleotides
    11.
    发明申请
    Methods for Amplification of Nucleic Acids Utilizing Clamp Oligonucleotides 审中-公开
    使用钳夹寡核苷酸扩增核酸的方法

    公开(公告)号：US20160130623A1

    公开(公告)日：2016-05-12

    申请号：US14773365

    申请日：2014-03-14

    申请人： Lyle J. ARNOLD ,  Norman C. NELSON

    发明人： Lyle J. Arnold

    IPC分类号： C12P19/34

    CPC分类号： C12P19/34 ,  C12Q1/6844 ,  C12Q2521/501 ,  C12Q2525/307 ,  C12Q2549/126

    摘要： The present invention provides methods of amplifying a target nucleic acid utilizing a clamp oligonucleotide comprising a first target-binding region on the 3′-terminus and a second target-binding region on the 5′-terminus and tether region in between. The tether region may comprise a variety of user-defined sequences or elements that allow for further manipulation of the target nucleic acid. Such as, for example, capture followed by amplification, identification and/or sequencing. The target-binding regions bind to the target nucleic acid, the 3′-terminus functions as a primer to initiate extension across the target nucleic acid sequence and ligation of the gap results in formation of a circularized nucleic acid. This circular template can be used in a variety of processes, including amplification and sequencing.

    摘要翻译： 本发明提供了使用包含3'末端的第一靶结合区和5'末端之间的第二靶结合区和其间的系链区的钳位寡核苷酸来扩增靶核酸的方法。 系链区域可以包含允许进一步操纵靶核酸的各种用户定义的序列或元件。 例如，进行捕获，随后进行扩增，鉴定和/或测序。 靶结合区域与靶核酸结合，3'-末端用作引物以引发靶核酸序列的延伸，并且间隙的连接导致形成环化的核酸。 该圆形模板可用于各种过程，包括扩增和测序。

    Methods for amplification of nucleic acids utilizing hairpin loop or duplex primers
    12.
    发明授权
    Methods for amplification of nucleic acids utilizing hairpin loop or duplex primers 有权
    使用发夹环或双链体引物扩增核酸的方法

    公开(公告)号：US09428747B2

    公开(公告)日：2016-08-30

    申请号：US14214621

    申请日：2014-03-14

    申请人： Lyle J. Arnold

    发明人： Lyle J. Arnold

    IPC分类号： C12P19/34 ,  C12N15/10 ,  C12Q1/68

    CPC分类号： C12Q1/6853 ,  C12N15/1065 ,  C12Q2521/501 ,  C12Q2525/301 ,  C12Q2563/185

    摘要： The present invention provides methods of amplifying a target nucleic acid utilizing duplex primer. The first strand of the primer comprises a random nucleotide sequence of about 6 to about 9 nucleotides in length that is able to hybridize to the target nucleic acid and a tag sequence. The second strand of the primer comprises a sequence complementary to the tag sequence allowing the primer to form a duplex and the ability to bind the tag sequence of the product nucleic acid for further amplification. The resulting nucleic acid produced contains tag sequences on both the 3′- and 5′-termini.

    摘要翻译： 本发明提供了利用双链体引物扩增靶核酸的方法。 引物的第一条链包含长度为约6至约9个核苷酸的随机核苷酸序列，其能够与靶核酸和标签序列杂交。 引物的第二条链包含与标签序列互补的序列，使得引物形成双链体并结合产物核酸的标签序列用于进一步扩增的能力。 产生的所得核酸含有3'和5'末端的标签序列。

    Methods for Amplifying a Complete Genome or Transcriptome
    13.
    发明申请
    Methods for Amplifying a Complete Genome or Transcriptome 审中-公开
    扩增完整基因组或转录组的方法

    公开(公告)号：US20140274811A1

    公开(公告)日：2014-09-18

    申请号：US14214634

    申请日：2014-03-14

    申请人： Lyle J. Arnold

    发明人： Lyle J. Arnold

    IPC分类号： C12N15/10

    CPC分类号： C12N15/1065

    摘要： The present invention provides methods for amplifying a complete genome or transcriptome. The genome or transcriptome is prepared as a set of target nucleic acids and mixed with a first primer. The first primer comprises a target-binding region having a first random sequence of about 6 to about 9 nucleotides and a tag sequence that contains one or more non-natural nucleotides. The first primer is annealed to the target nucleic acids and extended by polymerase to produce a first duplex nucleic acid. The target nucleic acid is removed from the first nucleic acid. A second primer is annealed to the first nucleic acid having a second random sequence of about 6 to about 9 nucleotides and a tag sequence that contains one or more non-natural nucleotides. The second primer is extended by polymerase to produce a second duplex nucleic acid. The second nucleic acid contains a tag sequence on one terminus and a complement of the tag sequence on the other. The first nucleic acid is removed from the second nucleic acid. A third primer is annealed to the second nucleic acid having the same sequence as the tag sequence or a portion thereof and at least one of the non-natural nucleotides of the tag sequence. The third primer is extended by polymerase to produce a third duplex nucleic acid. The second nucleic acid is removed from the third nucleic acid. The third primer is annealed to the second nucleic acid and the third nucleic acid. The third primer is extended by polymerase. Repeating these last three steps thereby results in amplification of the genome or transcriptome.

    摘要翻译： 本发明提供了扩增完整基因组或转录组的方法。 将基因组或转录组制备为一组靶核酸并与第一引物混合。 第一引物包含具有约6至约9个核苷酸的第一随机序列的靶结合区和含有一个或多个非天然核苷酸的标签序列。 将第一引物与靶核酸退火并通过聚合酶延伸以产生第一双链体核酸。 从第一核酸中除去靶核酸。 将第二引物退火至具有约6至约9个核苷酸的第二随机序列的第一核酸和含有一个或多个非天然核苷酸的标签序列。 通过聚合酶扩增第二引物以产生第二双链体核酸。 第二核酸在一个末端含有标签序列，另一个包含标签序列的互补序列。 从第二核酸中除去第一核酸。 第三引物与具有与标签序列或其部分相同序列的第二核酸和标签序列的至少一个非天然核苷酸退火。 通过聚合酶扩增第三引物以产生第三个双链体核酸。 从第三核酸中除去第二核酸。 将第三引物与第二核酸和第三核酸退火。 第三个引物通过聚合酶扩增。 重复这三个步骤，从而导致基因组或转录组的扩增。

    Methods for amplification of nucleic acids utilizing clamp oligonucleotides
    14.
    发明授权
    Methods for amplification of nucleic acids utilizing clamp oligonucleotides 有权

    公开(公告)号：US10995355B2

    公开(公告)日：2021-05-04

    申请号：US16231857

    申请日：2018-12-24

    申请人： Lyle J. Arnold

    发明人： Lyle J. Arnold

    IPC分类号： C12Q1/68 ,  C12P19/34 ,  G05D27/02 ,  H05B47/175 ,  C12Q1/6844

    摘要： The present invention provides methods of amplifying a target nucleic acid utilizing a clamp oligonucleotide comprising a first target-binding region on the 3′-terminus and a second target-binding region on the 5′-terminus and tether region in between. The tether region may comprise a variety of user-defined sequences or elements that allow for further manipulation of the target nucleic acid. Such as, for example, capture followed by amplification, identification and/or sequencing. The target-binding regions bind to the target nucleic acid, the 3′-terminus functions as a primer to initiate extension across the target nucleic acid sequence and ligation of the gap results in formation of a circularized nucleic acid. This circular template can be used in a variety of processes, including amplification and sequencing.

    Methods for Immobilizing Target Nucleic Acids Utilizing Combinatorial Capture Probes
    17.
    发明申请
    Methods for Immobilizing Target Nucleic Acids Utilizing Combinatorial Capture Probes 审中-公开

    公开(公告)号：US20180010173A1

    公开(公告)日：2018-01-11

    申请号：US15652372

    申请日：2017-07-18

    申请人： Lyle J. Arnold

    发明人： Lyle J. Arnold

    IPC分类号： C12Q1/68

    CPC分类号： C12Q1/6834 ,  C12Q2525/161 ,  C12Q2525/30 ,  C12Q2537/162

    摘要： The present invention provides methods for immobilizing target nucleic acids on a solid support utilizing combinatorial capture probe pairs. These pairs contain first and second capture oligonucleotides that each comprise a target binding region, a capture region and a stem region positioned between the target binding and capture regions. The target binding regions comprise nucleic acid sequences that allow them to hybridize to adjacent regions on the target nucleic acid. The stem regions have nucleic acid sequences that are complementary to each other and the capture regions each comprise a sequence that when positioned adjacent to one another produce a combined nucleic acid sequence that is complementary to a portion of an oligonucleotide bound to a solid support. When the first and second capture oligonucleotides are annealed to the target nucleic acid, the stem regions are brought together allowing them to hybridize, which in turn brings the capture regions together to produce a combined nucleic acid sequence. This combined nucleic acid sequence is then able to hybridize to the oligonucleotide bound to the solid support thereby immobilizing the target nucleic acid.

    Methods for amplification of nucleic acids utilizing hairpin loop or duplex primers
    18.
    发明申请
    Methods for amplification of nucleic acids utilizing hairpin loop or duplex primers 审中-公开
    使用发夹环或双链体引物扩增核酸的方法

    公开(公告)号：US20170009282A1

    公开(公告)日：2017-01-12

    申请号：US14999978

    申请日：2016-07-21

    申请人： Lyle J. Arnold

    发明人： Lyle J. Arnold

    IPC分类号： C12Q1/68

    CPC分类号： C12Q1/6853 ,  C12N15/1065 ,  C12Q2521/501 ,  C12Q2525/301 ,  C12Q2563/185

    摘要： The present invention provides methods of amplifying a target nucleic acid utilizing duplex primer. The first strand of the primer comprises a random nucleotide sequence of about 6 to about 9 nucleotides in length that is able to hybridize to the target nucleic acid and a tag sequence. The second strand of the primer comprises a sequence complementary to the tag sequence allowing the primer to form a duplex and the ability to bind the tag sequence of the product nucleic acid for further amplification. The resulting nucleic acid produced contains tag sequences on both the 3′- and 5′-termini.

    摘要翻译： 本发明提供了利用双链体引物扩增靶核酸的方法。 引物的第一条链包含长度为约6至约9个核苷酸的随机核苷酸序列，其能够与靶核酸和标签序列杂交。 引物的第二条链包含与标签序列互补的序列，使得引物形成双链体并结合产物核酸的标签序列用于进一步扩增的能力。 产生的所得核酸含有3'和5'末端的标签序列。

    Methods for Amplification of Nucleic Acids Utilizing Clamp Oligonucleotides
    19.
    发明申请
    Methods for Amplification of Nucleic Acids Utilizing Clamp Oligonucleotides 审中-公开

    公开(公告)号：US20190177760A1

    公开(公告)日：2019-06-13

    申请号：US16231857

    申请日：2018-12-24

    申请人： Lyle J. Arnold

    发明人： Lyle J. Arnold

    IPC分类号： C12P19/34 ,  C12Q1/6844

    摘要： The present invention provides methods of amplifying a target nucleic acid utilizing a clamp oligonucleotide comprising a first target-binding region on the 3′-terminus and a second target-binding region on the 5′-terminus and tether region in between. The tether region may comprise a variety of user-defined sequences or elements that allow for further manipulation of the target nucleic acid. Such as, for example, capture followed by amplification, identification and/or sequencing. The target-binding regions bind to the target nucleic acid, the 3′-terminus functions as a primer to initiate extension across the target nucleic acid sequence and ligation of the gap results in formation of a circularized nucleic acid. This circular template can be used in a variety of processes, including amplification and sequencing.

    Methods for amplification of nucleic acids utilizing hairpin loop or duplex primers
    20.
    发明授权
    Methods for amplification of nucleic acids utilizing hairpin loop or duplex primers 有权

    公开(公告)号：US10066262B2

    公开(公告)日：2018-09-04

    申请号：US14999978

    申请日：2016-07-21

    申请人： Lyle J. Arnold

    发明人： Lyle J. Arnold

    IPC分类号： C12P19/34 ,  C12Q1/6853 ,  C12N15/10

    摘要： The present invention provides methods of amplifying a target nucleic acid utilizing duplex primer. The first strand of the primer comprises a random nucleotide sequence of about 6 to about 9 nucleotides in length that is able to hybridize to the target nucleic acid and a tag sequence. The second strand of the primer comprises a sequence complementary to the tag sequence allowing the primer to form a duplex and the ability to bind the tag sequence of the product nucleic acid for further amplification. The resulting nucleic acid produced contains tag sequences on both the 3′- and 5′-termini.