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    • 3. 发明申请
    • Methods and compositions for amplification of genomic DNA
    • 用于扩增基因组DNA的方法和组合物
    • US20050147975A1
    • 2005-07-07
    • US10746871
    • 2003-12-24
    • Carol SchembriLeslie Leonard
    • Carol SchembriLeslie Leonard
    • C12P19/34C12Q1/68
    • C12Q1/6865
    • Methods for amplifying RNA from genomic DNA are provided. In the subject methods, a promoter-primer having a primer domain linked to an RNA polymerase promoter domain is first annealed to genomic DNA. The primer domain of the resultant annealed promoter-primer/genomic DNA complex is then extended to produce a double-stranded DNA molecule that has an RNA polymerase promoter domain. The resultant double-stranded DNA molecule is then transcribed into RNA product, e.g., labeled RNA product, using an RNA polymerase that is able to transcribe through the gap between the 5′ terminus of the promoter domain and the 3′ side of the genomic template. The subject methods find use a variety of different applications in which the preparation of amplified amounts of RNA from a genomic template is desired, where the amplification may be linear or geometric and may amplify the entire genome or only a select portion thereof. Also provided are kits for practicing the subject methods.
    • 提供了从基因组DNA扩增RNA的方法。 在本方法中,首先将与RNA聚合酶启动子结构域连接的引物结构域的启动子 - 引物退火至基因组DNA。 然后将所得退火的启动子 - 引物/基因组DNA复合物的引物结构域延伸以产生具有RNA聚合酶启动子结构域的双链DNA分子。 然后将得到的双链DNA分子转录成RNA产物,例如标记的RNA产物,使用能够通过启动子结构域的5'末端和基因组模板的3'侧之间的间隙转录的RNA聚合酶 。 本发明方法发现使用各种不同的应用,其中需要从基因组模板制备扩增量的RNA,其中扩增可以是线性的或几何的,并且可以扩增整个基因组或仅扩增其选择部分。 还提供了用于实践主题方法的工具包。