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    TRANSPOSON END COMPOSITIONS AND METHODS FOR MODIFYING NUCLEIC ACIDS
    2.
    发明申请
    TRANSPOSON END COMPOSITIONS AND METHODS FOR MODIFYING NUCLEIC ACIDS 有权
    TRANSPOSON END COMPOSITIONS AND METHODS FOR MODIZING NUCLEIC ACIDS

    公开(公告)号：US20140162897A1

    公开(公告)日：2014-06-12

    申请号：US14148463

    申请日：2014-01-06

    申请人： Haiying Li Grunenwald ,  Nicholas Caruccio ,  Jerome Jendrisak ,  Gary Dahl

    发明人： Haiying Li Grunenwald ,  Nicholas Caruccio ,  Jerome Jendrisak ,  Gary Dahl

    IPC分类号： C12Q1/68

    CPC分类号： C12N15/1065 ,  C12N15/10 ,  C12N15/1093 ,  C12P19/34 ,  C12Q1/6806 ,  C12Q1/6855 ,  C12Q1/6869 ,  C12Q1/6874 ,  C12Q2535/122 ,  C12Q2521/507

    摘要： The present invention provides methods, compositions and kits for using a transposase and a transposon end for generating extensive fragmentation and 5′-tagging of double-stranded target DNA in vitro, then using a DNA polymerase for generating 5′- and 3′-tagged single-stranded DNA fragments without performing a PCR amplification reaction, wherein the first tag on the 5′-ends exhibits the sequence of the transferred transposon end and optionally, an additional arbitrary sequence, and the second tag on the 3′-ends exhibits a different sequence from the sequence exhibited by the first tag. The method is useful for generating 5′- and 3′-tagged DNA fragments for use in a variety of processes, including processes for metagenomic analysis of DNA in environmental samples, copy number variation (CNV) analysis of DNA, and comparative genomic sequencing (CGS), including massively parallel DNA sequencing (so-called “next-generation sequencing.)

    摘要翻译： 本发明提供了使用转座酶和转座子末端在体外产生双链靶DNA的广泛断裂和5'标记的方法，组合物和试剂盒，然后使用DNA聚合酶产生5'-和3'标记 单链DNA片段，而不进行PCR扩增反应，其中5'-末端的第一个标签显示转移的转座子末端的序列和任选的另外的任意序列，3'-末端的第二个标签显示出 与第一个标签展示的序列不同的序列。 该方法可用于产生用于各种过程的5'-和3'标记的DNA片段，包括环境样品中DNA的宏基因组分析过程，DNA拷贝数变异（CNV）分析和比较基因组测序（ CGS），包括大规模并行DNA测序（所谓的“下一代测序”）。

    Selective 5′ ligation tagging of RNA
    3.
    发明授权
    Selective 5′ ligation tagging of RNA 有权
    RNA的选择性5'连接标记

    公开(公告)号：US08163491B2

    公开(公告)日：2012-04-24

    申请号：US12707243

    申请日：2010-02-17

    申请人： Jerome Jendrisak ,  Ramesh Vaidyanathan ,  Gary Dahl

    发明人： Jerome Jendrisak ,  Ramesh Vaidyanathan ,  Gary Dahl

    IPC分类号： C12Q1/68

    CPC分类号： C12Q1/6806 ,  C12N15/1096 ,  C12Q1/6855 ,  C12Q2525/207 ,  C12Q2525/125 ,  C12Q2521/525

    摘要： The present invention provides novel compositions, kits and methods employing RNA 5′ polyphosphatases, RNA 5′ monophosphatases, capping enzymes, decapping enzymes, nucleic acid pyrophosphatases and RNA ligases, as well as other enzymes, for selective 5′ ligation tagging of desired classes of RNA molecules that differ with respect to particular chemical moieties on their 5′ ends. The 5′tagged RNA molecules can be used for synthesis of tagged first-stand cDNA, double-stranded cDNA, and sense or antisense RNA for a variety of uses.

    摘要翻译： 本发明提供使用RNA 5'多磷酸酶，RNA 5'单磷酸酶，封端酶，脱蛋白酶，核酸焦磷酸酶和RNA连接酶以及其它酶的新型组合物，试剂盒和方法，用于选择性5'连接标记所需类别的 相对于其5'端的特定化学部分不同的RNA分子。 5'标记的RNA分子可用于合成标记的第一支cDNA，双链cDNA和用于各种用途的正义或反义RNA。

    Synthesis of tagged nucleic acids
    4.
    发明授权
    Synthesis of tagged nucleic acids 有权
    标记核酸的合成

    公开(公告)号：US08039214B2

    公开(公告)日：2011-10-18

    申请号：US12165324

    申请日：2008-06-30

    申请人： Gary Dahl ,  Roy Rabindranauth Sooknanan

    发明人： Gary Dahl ,  Roy Rabindranauth Sooknanan

    IPC分类号： C12Q1/68 ,  C12P19/34

    CPC分类号： C12N15/1096 ,  C12Q1/6865 ,  C12Q2525/186 ,  C12Q2525/155

    摘要： The present invention relates generally to methods, compositions and kits for synthesizing sense RNA molecules from one or more RNA molecules of interest in a sample. In exemplary embodiments, the methods use a terminal tagging oligoribonucleotide (rTTO) to join a DNA sequence tag to the 3′-termini of first-strand cDNA molecules. The use of an rTTO comprising ribonucleotides results in decreased oligonucleotide-derived background synthesis of RNA in the absence of sample RNA and, surprisingly and unexpectedly, also results in significantly increased yields of sense RNA molecules that exhibit sequences that are substantially identical to those of the RNA molecules of interest in the sample. The sense RNA molecules also have an RNA sequence tag on their 5′-termini that is useful for fixing the lengths of sense RNA molecules that are synthesized in a second or subsequent round.

    摘要翻译： 本发明一般涉及从样品中一个或多个感兴趣的RNA分子合成有义RNA分子的方法，组合物和试剂盒。 在示例性实施方案中，所述方法使用末端标记寡核糖核苷酸（rTTO）将DNA序列标签连接到第一链cDNA分子的3'-末端。 包含核糖核苷酸的rTTO的使用导致在不存在样品RNA的情况下寡核苷酸衍生的RNA背景合成减少，令人惊讶地和出人意料地也导致显着提高正义RNA分子的产量，其显示出与 样品中感兴趣的RNA分子。 有义RNA分子在其5'-末端也具有RNA序列标签，其可用于固定在第二次或随后的一轮中合成的有义RNA分子的长度。

    TRANSPOSON END COMPOSITIONS AND METHODS FOR MODIFYING NUCLEIC ACIDS
    5.
    发明申请
    TRANSPOSON END COMPOSITIONS AND METHODS FOR MODIFYING NUCLEIC ACIDS 有权
    TRANSPOSON END COMPOSITIONS AND METHODS FOR MODIZING NUCLEIC ACIDS

    公开(公告)号：US20100120098A1

    公开(公告)日：2010-05-13

    申请号：US12605337

    申请日：2009-10-24

    申请人： Haiying Li Grunenwald ,  Nicholas Caruccio ,  Jerome Jendrisak ,  Gary Dahl

    发明人： Haiying Li Grunenwald ,  Nicholas Caruccio ,  Jerome Jendrisak ,  Gary Dahl

    IPC分类号： C12P19/34 ,  C07H21/00 ,  C12N9/00 ,  C12N9/10

    CPC分类号： C12N15/1065 ,  C12N15/10 ,  C12N15/1093 ,  C12P19/34 ,  C12Q1/6806 ,  C12Q1/6855 ,  C12Q1/6869 ,  C12Q1/6874 ,  C12Q2535/122 ,  C12Q2521/507

    摘要： The present invention provides methods, compositions and kits for using a transposase and a transposon end for generating extensive fragmentation and 5′-tagging of double-stranded target DNA in vitro, then using a DNA polymerase for generating 5′- and 3′-tagged single-stranded DNA fragments without performing a PCR amplification reaction, wherein the first tag on the 5′-ends exhibits the sequence of the transferred transposon end and optionally, an additional arbitrary sequence, and the second tag on the 3′-ends exhibits a different sequence from the sequence exhibited by the first tag. The method is useful for generating 5′- and 3′-tagged DNA fragments for use in a variety of processes, including processes for metagenomic analysis of DNA in environmental samples, copy number variation (CNV) analysis of DNA, and comparative genomic sequencing (CGS), including massively parallel DNA sequencing (so-called “next-generation sequencing.)

    摘要翻译： 本发明提供了使用转座酶和转座子末端在体外产生双链靶DNA的广泛断裂和5'标记的方法，组合物和试剂盒，然后使用DNA聚合酶产生5'-和3'标记 单链DNA片段，而不进行PCR扩增反应，其中5'-末端的第一个标签显示转移的转座子末端的序列和任选的另外的任意序列，3'-末端的第二个标签显示出 与第一个标签展示的序列不同的序列。 该方法可用于产生用于各种过程的5'-和3'标记的DNA片段，包括环境样品中DNA的宏基因组分析过程，DNA拷贝数变异（CNV）分析和比较基因组测序（ CGS），包括大规模并行DNA测序（所谓的“下一代测序”）。

    Synthesis of tagged nucleic acids
    9.
    发明授权
    Synthesis of tagged nucleic acids 有权
    标记核酸的合成

    公开(公告)号：US08329887B2

    公开(公告)日：2012-12-11

    申请号：US13237451

    申请日：2011-09-20

    申请人： Gary Dahl ,  Roy Rabindranauth Sooknanan

    发明人： Gary Dahl ,  Roy Rabindranauth Sooknanan

    IPC分类号： C07H21/04 ,  C12P19/34

    CPC分类号： C12N15/1096 ,  C12Q1/6865 ,  C12Q2525/186 ,  C12Q2525/155

    摘要： The present invention relates generally to methods, compositions and kits for synthesizing sense RNA molecules from one or more RNA molecules of interest in a sample. In exemplary embodiments, the methods use a terminal tagging oligoribonucleotide (rTTO) to join a DNA sequence tag to the 3′-termini of first-strand cDNA molecules. The use of an rTTO comprising ribonucleotides results in decreased oligonucleotide-derived background synthesis of RNA in the absence of sample RNA and, surprisingly and unexpectedly, also results in significantly increased yields of sense RNA molecules that exhibit sequences that are substantially identical to those of the RNA molecules of interest in the sample. The sense RNA molecules also have an RNA sequence tag on their 5′-termini that is useful for fixing the lengths of sense RNA molecules that are synthesized in a second or subsequent round.

    摘要翻译： 本发明一般涉及从样品中一个或多个感兴趣的RNA分子合成有义RNA分子的方法，组合物和试剂盒。 在示例性实施方案中，所述方法使用末端标记寡核糖核苷酸（rTTO）将DNA序列标签连接到第一链cDNA分子的3'-末端。 包含核糖核苷酸的rTTO的使用导致在不存在样品RNA的情况下寡核苷酸衍生的RNA背景合成减少，令人惊讶地和出人意料地也导致显着提高正义RNA分子的产量，其显示出与 样品中感兴趣的RNA分子。 有义RNA分子在其5'-末端也具有RNA序列标签，其可用于固定在第二次或随后的一轮中合成的有义RNA分子的长度。