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    • 4. 发明申请
    • METHODS REACTION MIXTURES AND KITS FOR LIGATING POLYNUCLEOTIDES
    • 方法反应混合物和用于引发多核苷酸的试剂盒
    • WO2006005055A3
    • 2006-03-16
    • PCT/US2005023737
    • 2005-06-30
    • APPLERA CORPANDERSEN MARKWENZ MICHAEL HCHEN CAIFUKARGER ACHIM
    • ANDERSEN MARKWENZ MICHAEL HCHEN CAIFUKARGER ACHIM
    • C12Q1/48C12Q1/527C12Q1/68
    • C12Q1/6855C12P19/34
    • The present teachings pertain to methods, reaction mixtures, and kits for ligating polynucleotides. In some embodiments, a heat-activatable ligation agent, a phosphorylation agent, and a decontamination agent are included in the same ligation reaction mixture with at least one probe set, at least one linker set, and at least one target polynucleotide. A reaction at a first temperature results in hybridization of the probes to the target, phosphorylation of the probes, and decontamination of unwanted reaction components. A reaction at a second temperature results in the ligation of the probes together. In some embodiments, the present teachings are applied in highly multi-plexed ligation reactions in which a plurality of single nucleotide polymorphisms in a plurality of target polynucleotides are queried, and eventually detected using a mobility dependent analysis technique.
    • 本教导涉及用于连接多核苷酸的方法,反应混合物和试剂盒。 在一些实施方案中,与至少一个探针组,至少一个接头组和至少一个靶多核苷酸在相同的连接反应混合物中包括可热激活的连接剂,磷酸化剂和去污剂。 在第一温度下的反应导致探针与靶的杂交,探针的磷酸化,以及不期望的反应成分的去污。 在第二温度下的反应导致探头连接在一起。 在一些实施方案中,本教导应用于高多重连接的连接反应中,其中查询多个目标多核苷酸中的多个单核苷酸多态性,并且最终使用迁移率依赖性分析技术检测。