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    NUCLEIC ACID MONOMERS WITH 2'-CHEMICAL MOIETIES
    3.
    发明申请
    NUCLEIC ACID MONOMERS WITH 2'-CHEMICAL MOIETIES 审中-公开
    具有2个化学成分的核酸单体

    公开(公告)号：US20070218490A1

    公开(公告)日：2007-09-20

    申请号：US11686894

    申请日：2007-03-15

    申请人： Andrei Laikhter ,  Joseph Walder ,  Mark Behlke

    发明人： Andrei Laikhter ,  Joseph Walder ,  Mark Behlke

    IPC分类号： C12Q1/68 ,  C12P19/34 ,  C07H21/04

    CPC分类号： C12P19/34 ,  C07H21/04

    摘要： The invention provides nucleic acid monomers with a 2′-modification that are useful for the incorporation of dyes or blocking groups. The monomers can be incorporated on the 3′-end of a dual labeled probe to inhibit PCR polymerase extension during PCR. The polymerase is inhibited from extending the probe at the 3′-hydroxyl group when the monomer is present; there is no need to add a chemical moiety to the 3′-hydroxyl or remove the 3′-hydroxyl. The monomers can also be incorporated internally or at the 5′-end of the oligonucleotide. A detectable label, such as a fluorescent or quenching dye, can be incorporated on the 2′-position of such monomers.

    摘要翻译： 本发明提供具有2'-修饰的核酸单体，其可用于引入染料或封闭基团。 单体可以掺入双标记探针的3'末端，以抑制PCR聚合酶扩增。 当存在单体时，聚合酶被抑制在3'-羟基延伸探针; 不需要向3'-羟基添加化学部分或除去3'-羟基。 单体也可以在内部或寡核苷酸的5'-末端引入。 可检测的标记，例如荧光或淬灭染料，可以掺入这些单体的2'-位上。

    Methods for amplifying polymeric nucleic acids
    5.
    发明申请
    Methods for amplifying polymeric nucleic acids 有权
    扩增聚合核酸的方法

    公开(公告)号：US20080038724A9

    公开(公告)日：2008-02-14

    申请号：US10911652

    申请日：2004-08-02

    申请人： Mark Behlke ,  Joseph Walder ,  Jeffrey Manthey

    发明人： Mark Behlke ,  Joseph Walder ,  Jeffrey Manthey

    IPC分类号： C12Q1/68 ,  C12P19/34

    CPC分类号： C12Q1/686 ,  C12Q1/6848 ,  C12Q1/6853 ,  C12Q2527/143 ,  C12Q2525/101 ,  C12Q2549/119 ,  C12Q2525/186

    摘要： The invention provides compositions and methods for amplifying nucleic acid polymer sequences in a high complexity nucleic acid sample. The unique compositions of the invention include a primer set composed of a mixture of two types of primers for DNA synthesis. For extension in one direction, the primers all contain modifications that destroy their ability to serve as templates that can be copied by DNA polymerases. For extension in the opposite direction the set includes at least one primer that can serve as a template and be replicated by DNA polymerases throughout its length. The method can be carried out by mixing the nucleic acid polymer sequence of interest with the set of DNA synthesis primers in an amplification reaction mixture. The reaction mixture is then subjected to temperature cycling analogous to the temperature cycling in PCR reactions. At least one primer in the primer set hybridizes to the nucleic acid polymer. It is preferred that the non-replicable primer hybridizes to the nucleic acid polymer and is extended to produce an extension product that contains sequence from the nucleic acid polymer to which the replicable primer then hybridizes. Of course, if the nucleic acid polymer is double stranded, both the replicable and nonreplicable primers will hybridize and be extended by DNA polymerase.

    摘要翻译： 本发明提供用于在高复杂度核酸样品中扩增核酸聚合物序列的组合物和方法。 本发明的独特组合物包括由两种用于DNA合成的引物的混合物组成的引物组。 为了在一个方向延伸，引物都包含破坏其作为可以由DNA聚合酶复制的模板的能力的修饰。 为了在相反方向延伸，该组包括至少一个引物，其可以用作模板并且在其整个长度上由DNA聚合酶复制。 该方法可以通过在扩增反应混合物中混合目的核酸聚合物序列与DNA合成引物组来进行。 然后将反应混合物进行类似于PCR反应中的温度循环的温度循环。 引物组中的至少一个引物与核酸聚合物杂交。 优选地，不可复制的引物与核酸聚合物杂交并延伸以产生包含来自核酸聚合物的序列的延伸产物，然后可复制的引物随后杂交。 当然，如果核酸聚合物是双链的，则可复制和不可复制的引物都将与DNA聚合酶杂交并延伸。