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    FINE-TUNED ULTRASPECIFIC NUCLEIC ACID HYBRIDIZATION PROBES
    3.
    发明公开
    FINE-TUNED ULTRASPECIFIC NUCLEIC ACID HYBRIDIZATION PROBES 审中-公开
    FEINABGESTIMMTE ULTRASPEZIFISCHENUKLEINSÄURE-HYBRIDISIERUNGSSONDEN

    公开(公告)号：EP3084051A1

    公开(公告)日：2016-10-26

    申请号：EP14873077.3

    申请日：2014-08-27

    申请人： William Marsh Rice University

    发明人： ZHANG, David, Yu ,  WANG, Juexiano ,  WU, Ruojia

    IPC分类号： C40B30/04 ,  C07H21/04 ,  C12Q1/68

    摘要： Compositions and methods for highly specific nucleic acid probes and primers are provided. The probe system comprises a complement strand and a protector stand that form a partially double-stranded probe. The reaction standard free energy of hybridization between the probe and target nucleic acid as determined by Expression 1 (ΔG°rxn=ΔG°t-TC−ΔG°nh-PC+(ΔG°v-TC−ΔG°h-PC)) is from about −4 kcal/mol to about +4 kcal/mol. Alternatively, the reaction standard free energy of hybridization between the probe and target nucleic acid is determined by Expression 1 to be within 5 kcal/mol of the standard free energy as determined by Expression 2 (−Rτ ln(([P]0−[C]0)/[C]0)]), where the [P]0 term of Expression 2 equals the concentration of the protector strand and the [C]0 term of Expression 2 equals the concentration of the complement strand. In addition, a method for on-the-fly fine tuning of a reaction using the present probe is provided.

    摘要翻译： 提供了高度特异性核酸探针和引物的组合物和方法。 探针系统包括形成部分双链探针的补体链和保护架。 通过表达式1（ΔG°rxn =ΔG°t-TC-ΔG°nh-PC +（ΔG°v-TC-ΔG°h-PC））测定的探针和靶核酸之间的反应标准自由能为 约-4kcal / mol至约+4kcal / mol。 或者，探针和靶核酸之间的反应标准自由能通过表达式1确定为通过表达式2确定的标准自由能的5kcal / mol（-Rτln（（[P] O- [ C] 0）/ [C] 0）]），其中表达式2的[P] 0项等于保护基团的浓度，并且表达式2的[C] 0项等于补体链的浓度。 另外，提供了使用本探针进行的使反应动态微调的方法。