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9 条记录 (耗时 0.052 秒)

    Method of nucleic acid cassette assembly
    8.
    发明授权
    Method of nucleic acid cassette assembly 有权

    公开(公告)号：US10041060B2

    公开(公告)日：2018-08-07

    申请号：US11635355

    申请日：2006-12-06

    申请人： J. Craig Venter ,  Hamilton O. Smith ,  Clyde A. Hutchison, III ,  Daniel G. Gibson

    发明人： J. Craig Venter ,  Hamilton O. Smith ,  Clyde A. Hutchison, III ,  Daniel G. Gibson

    IPC分类号： C12Q1/68 ,  C12N15/10 ,  C12N15/66 ,  C07H21/02

    摘要： Methods are provided for constructing a synthetic genome, comprising generating and assembling nucleic acid cassettes comprising portions of the genome, wherein at least one of the nucleic acid cassettes is constructed from nucleic acid components that have been chemically synthesized, or from copies of the chemically synthesized nucleic acid components. In one embodiment, the entire synthetic genome is constructed from nucleic acid components that have been chemically synthesized, or from copies of the chemically synthesized nucleic acid components. Rational methods may be used to design the synthetic genome (e.g., to establish a minimal genome and/or to optimize the function of genes within a genome, such as by mutating or rearranging the order of the genes). Synthetic genomes of the invention may be introduced into vesicles (e.g., bacterial cells from which part or all of the resident genome has been removed, or synthetic vesicles) to generate synthetic cells. Synthetic genomes or synthetic cells may be used for a variety of purposes, including the generation of synthetic fuels, such as hydrogen or ethanol.

    Amplification and cloning of single DNA molecules using rolling circle amplification
    9.
    发明授权
    Amplification and cloning of single DNA molecules using rolling circle amplification 有权
    使用滚环扩增扩增和克隆单个DNA分子

    公开(公告)号：US08497069B2

    公开(公告)日：2013-07-30

    申请号：US11919515

    申请日：2006-05-01

    申请人： Clyde A. Hutchison, III ,  Hamilton O. Smith

    发明人： Clyde A. Hutchison, III ,  Hamilton O. Smith

    IPC分类号： C12Q1/68 ,  C12P19/34

    CPC分类号： C12N15/1075 ,  C12N15/64 ,  C12Q1/6848 ,  C12Q2531/119 ,  C12Q2527/143 ,  C12Q2525/204 ,  C12Q2531/125

    摘要： The present invention relates, e.g., to a method for amplifying a small number of copies (e.g. a single copy) of a single-stranded circular DNA molecule (e.g. having a size of about 5-6 kb) by an isothermal rolling circle mechanism, using random or partially random primers and a F29-type DNA polymerase. The method, which can also be used for amplifying DNAs by non-rolling types of multiple displacement amplification, comprises incubating the reaction components in a small volume, e.g. about 10 μl or less, such as about 0.6 μl or less. The degree of amplification can be about 109 fold, or higher. A method for cell-free cloning of DNA, using the rolling circle amplification method of the invention, is described.

    摘要翻译： 本发明涉及例如通过等温滚圆机制扩增单链环状DNA分子（例如大小约5-6kb）的少量拷贝（例如单拷贝）的方法， 使用随机或部分随机引物和F29型DNA聚合酶。 也可以用于通过非滚动类型的多位置扩增来扩增DNA的方法包括将反应组分以小体积例如温育。 约10个或更少，例如约0.6毫摩尔或更少。 扩增程度可以是约109倍或更高。 描述了使用本发明的滚环扩增方法进行无细胞克隆DNA的方法。