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1. 发明申请

WO2004044239A1 IMPROVED METHODS FOR GENERATING MULTIPLE RNA COPIES 审中-公开
标题翻译：用于产生多个RNA拷贝的改进方法
公开(公告)号：WO2004044239A1
公开(公告)日：2004-05-27
申请号：PCT/EP2003/012058
申请日：2003-10-30
申请人： PAMGENE B.V. , BOENDER, Piet
发明人： BOENDER, Piet
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6865 , C12Q2521/107 , C12Q2521/101 , C12Q2525/186 , C12Q2521/325
摘要： The present invention is directed to a novel method of efficiently synthesizing, in a non-specific manner, multiple copies of a target RNA. The present invention also relates to kits relating to the same and the use of these copies for determining gene expression pattern. In particular, the present invention relates to a method for generating multiple RNA copies comprising the steps of (a) providing a sample comprising target RNA; wherein said sample is simultaneously contacted with an oligonucleotide comprising at its 5' side a promoter sequence recognized by an RNA polymerase, wherein each oligonucleotide further comprises a target hybridising sequence, which is a random sequence or a predetermined sequence and possibly a modification at its 3' terminal end in such a way that extension therefrom is prohibited; and, an enzyme having DNA polymerase activity; an enzyme having RNase H activity; an enzyme having RNA polymerase activity; and, sufficient amounts of dNTPs and rNTPs; and, (b) maintaining the resulting reaction mixture under the appropriate conditions for a sufficient amount of time for the enzymatic processes to take place.
摘要翻译：本发明涉及以非特异性方式有效合成靶RNA的多个拷贝的新方法。本发明还涉及与其相关的试剂盒以及用于确定基因表达模式的这些拷贝的用途。特别地，本发明涉及产生多个RNA拷贝的方法，包括以下步骤：（a）提供包含靶RNA的样品; 其中所述样品与包含在其5'侧的包含RNA聚合酶识别的启动子序列的寡核苷酸同时接触，其中每个寡核苷酸还包含目标杂交序列，其是随机序列或预定序列，并且可能在其3 终端以这样的方式被禁止; 和具有DNA聚合酶活性的酶; 具有RNase H活性的酶; 具有RNA聚合酶活性的酶; 和足量的dNTPs和rNTPs; 和（b）将所得反应混合物在适当的条件下保持足够的时间以进行酶促处理。

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