Transposon end compositions and methods for modifying nucleic acids
    4.
    发明授权
    Transposon end compositions and methods for modifying nucleic acids 有权
    转座子末端组合物和修饰核酸的方法

    公开(公告)号:US09080211B2

    公开(公告)日:2015-07-14

    申请号:US12605337

    申请日:2009-10-24

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

    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”)。

    TRANSPOSON END COMPOSITIONS AND METHODS FOR MODIFYING NUCLEIC ACIDS
    6.
    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

    IPC分类号: C12Q1/68

    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”)。

    TRANSPOSON END COMPOSITIONS AND METHODS FOR MODIFYING NUCLEIC ACIDS
    7.
    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

    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”)。

    Compositions and methods employing 5' phosphate-dependent nucleic acid exonucleases
    10.
    发明申请
    Compositions and methods employing 5' phosphate-dependent nucleic acid exonucleases 审中-公开
    使用5'磷酸依赖性核酸外切核酸酶的组合物和方法

    公开(公告)号:US20060240451A1

    公开(公告)日:2006-10-26

    申请号:US11352172

    申请日:2006-02-09

    IPC分类号: C12Q1/68 C07H21/02 C12P19/34

    摘要: The present invention relates to compositions and methods employing 5′-phosphate-dependent nucleic acid exonucleases. In particular, the present invention provides kits and methods employing 5′-phosphate-dependent nucleic acid exonucleases for selective enrichment, isolation and amplification of a particular set of desired nucleic acid molecules from samples that also contain undesired nucleic acid molecules for a variety of uses. In preferred embodiments, the desired nucleic acid molecules comprise prokaryotic and/or eukaryotic mRNA.

    摘要翻译: 本发明涉及采用5'-磷酸依赖性核酸外切核酸酶的组合物和方法。 特别地,本发明提供了使用5'-磷酸依赖性核酸外切核酸酶用于选择性富集,分离和扩增特定的一组所需核酸分子的试剂盒和方法,该样品还含有用于各种用途的不想要的核酸分子的样品 。 在优选的实施方案中,所需的核酸分子包含原核和/或真核的mRNA。