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    Partially double-stranded nucleic acids, methods of making, and use thereof
    8.
    发明申请
    Partially double-stranded nucleic acids, methods of making, and use thereof 审中-公开
    部分双链核酸，制备方法和用途

    公开(公告)号：US20050095606A1

    公开(公告)日：2005-05-05

    申请号：US10757468

    申请日：2004-01-15

    申请人： Glenn Hoke ,  John Hartwell ,  Adam Steel

    发明人： Glenn Hoke ,  John Hartwell ,  Adam Steel

    IPC分类号： C12Q1/68 ,  C12P19/34

    CPC分类号： C12Q1/6844 ,  C12Q1/6853 ,  C12Q2525/113 ,  C12Q2521/319 ,  C12Q2521/531 ,  C12Q2525/161 ,  C12Q2523/319 ,  C12Q2523/107

    摘要： A process is disclosed for generating at least one partially double-stranded target nucleic acid, which contains at least one single-stranded region at a terminal end. The process comprises the steps of (a) providing at least one primer, P1, containing at least one labile nucleotide; (b) combining at least one target nucleic acid sequence with P1 to generate a double-stranded polynucleotide containing at least one labile nucleotide; (c) exposing the double-stranded polynucleotide to conditions that promote single-strand cleavage of the polynucleotide at the site of the at least one labile nucleotide of primer P1; and (d) exposing the cleaved polynucleotide to conditions that promote the dissociation of the cleaved portions of primer P1 from a terminal end. The labile nucleotide may be dUTP, wherein the single-stranded cleavage of the polynucleotide at the site of the labile nucleotide occurs by treatment with uracil N-glycosylase.

    摘要翻译： 公开了用于产生至少一个在末端含有至少一个单链区的部分双链靶核酸的方法。 该方法包括以下步骤：（a）提供至少一种含有至少一个不稳定核苷酸的引物P1; （b）将至少一个靶核酸序列与P1结合以产生含有至少一个不稳定核苷酸的双链多核苷酸; （c）将双链多核苷酸暴露于在引物P1的至少一个不稳定核苷酸的位点处促进单链切割多核苷酸的条件; 和（d）将切割的多核苷酸暴露于促进引物P1的切割部分从终末端解离的条件。 不稳定核苷酸可以是dUTP，其中在不稳定核苷酸位点处的多核苷酸的单链切割通过尿嘧啶N-糖基化酶的处理发生。