DNA amplification and sequencing using DNA molecules generated by random fragmentation
    5.
    发明授权
    DNA amplification and sequencing using DNA molecules generated by random fragmentation 有权
    DNA扩增和使用随机碎裂产生的DNA分子进行测序

    公开(公告)号:US08815504B2

    公开(公告)日:2014-08-26

    申请号:US12697886

    申请日:2010-02-01

    IPC分类号: C12Q1/68 C12P19/34

    摘要: The present invention is directed to methods to prepare a DNA molecule or a plurality of DNA molecules by random fragmentation. In some embodiments, the present invention regards preparing a template for DNA sequencing by random fragmentation. In specific embodiments, the random fragmentation comprises chemical fragmentation, mechanical fragmentation, or enzymatic fragmentation. In further specific embodiments, a universal sequence is attached to the 3′ end of the DNA fragments, such as by ligation of an adaptor sequence or by homopolymeric tailing with terminal deoxynucleotidyltransferase. In other embodiments, a library is prepared with methods of the present invention.

    摘要翻译: 本发明涉及通过随机碎裂制备DNA分子或多个DNA分子的方法。 在一些实施方案中,本发明涉及通过随机断裂制备用于DNA测序的模板。 在具体实施方案中,随机碎裂包括化学断裂,机械破碎或酶促碎裂。 在进一步的具体实施方案中,通用序列附着于DNA片段的3'末端,例如通过连接衔接子序列或通过与末端脱氧核苷酸转移酶的均聚尾料连接。 在其它实施方案中,使用本发明的方法制备文库。

    Isolation of CpG islands by thermal segregation and enzymatic selection-amplification method
    6.
    发明授权
    Isolation of CpG islands by thermal segregation and enzymatic selection-amplification method 有权
    通过热分离和酶选扩增方法分离CpG岛

    公开(公告)号:US08409804B2

    公开(公告)日:2013-04-02

    申请号:US11367046

    申请日:2006-03-02

    IPC分类号: C12Q1/68 C12P19/34

    摘要: The present invention concerns isolation, library preparation and selective amplification from a compositionally heterogeneous pool of DNA fragments of a fraction of molecules, such as those originating from promoter CpG islands and characterized by a high GC content. In particular, the process utilizes a heat-induced segregation of DNA molecules into GC-poor, single-stranded molecule fractions and GC-rich, double-stranded molecule fractions, with subsequent enzymatic conversion of the GC-rich, double-stranded DNA molecules into a library, and, optionally, amplification. In specific embodiments, the isolation process is used to generate promoter-enriched genomic and methylome libraries for research and diagnostic applications, for example.

    摘要翻译: 本发明涉及分离,文库制备和从部分分子的DNA片段的组成不均一聚集体(例如源自启动子CpG岛的那些DNA片段)的选择性扩增,并且特征在于高GC含量。 特别地,该方法利用热分解DNA分子到GC-不良的单链分子级分和富含GC的双链分子级分,随后富含GC的双链DNA分子的酶转化 进入文库,可选地,扩增。 在具体实施方案中,例如,分离过程用于产生用于研究和诊断应用的富含启动子的基因组和甲基化文库。

    Genome walking by selective amplification of nick-translate DNA library and amplification from complex mixtures of templates
    9.
    发明申请
    Genome walking by selective amplification of nick-translate DNA library and amplification from complex mixtures of templates 审中-公开
    通过选择性扩增切口翻译DNA文库并从模板的复杂混合物中扩增的基因组

    公开(公告)号:US20050032104A1

    公开(公告)日:2005-02-10

    申请号:US10909516

    申请日:2004-08-02

    IPC分类号: C12Q1/68 C12P19/34

    CPC分类号: C12Q1/6869 C12Q1/6876

    摘要: Improved methods and reagents for chromosome walking of nucleic acid are discussed herein. A library of amplifiable nick translation molecules is generated, and a chromosome walk is initiated from a known sequence in the nucleic acid by producing at least one nick translate molecule, sequencing part of the nick translate molecule, and producing a second nick translate molecule by initiating the primer extension from the region of the obtained sequence of the prior nick translate molecule.

    摘要翻译: 本文讨论了用于核酸染色体行走的改进方法和试剂。 产生可扩增切口平移分子的文库,通过产生至少一个切口平移分子,切割平移分子的测序部分,并通过引发产生第二切口平移分子,从核酸中的已知序列引发染色体步态 来自所获得的先前切口平移分子序列的区域的引物延伸。