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    • 4. 发明申请
    • Detection format for hot start real time polymerase chain reaction
    • 检测格式为热启动实时聚合酶链反应
    • US20090181401A1
    • 2009-07-16
    • US12407339
    • 2009-03-19
    • Dieter HeindlWaltraud AnkenbauerFrank Laue
    • Dieter HeindlWaltraud AnkenbauerFrank Laue
    • C12Q1/68
    • C12Q1/6818C12Q2561/113C12Q2531/113C12Q2521/319
    • The present invention is directed to a method and a composition for amplifying and detecting a target nucleic comprising subjecting said target nucleic acid to a real time PCR amplification reaction in the presence of a thermostable DNA polymerase, a thermostable double strand dependent 3′-5′ exonuclease having a temperature optimum above 37° C., a pair of amplification primers, deoxynucleoside triphosphates, a detecting oligonucleotide carrying a first label and a second label, said first label being capable of acting as a fluorescent reporter entity when excited with light of an appropriate wavelength, said second label being capable of acting as a fluorescence quenching entity of said fluorescent reporter entity, characterized in that one label is bound to the 3′ end of said detecting oligonucleotide, and further characterized in that the other label is bound either internally or at the 5′ end of said detecting oligonucleotide, and monitoring fluorescence of said fluorescent reporter entity at least after a plurality of amplification cycles.
    • 本发明涉及用于扩增和检测靶核酸的方法和组合物,其包括在热稳定性DNA聚合酶,耐热双链依赖性3'-5'的存在下使所述靶核酸进行实时PCR扩增反应, 温度最高在37℃以上的外切核酸酶,一对扩增引物,脱氧核苷三磷酸,携带第一标记和第二标记的检测寡核苷酸,所述第一标记当用 所述第二标记能够用作所述荧光报道实体的荧光猝灭实体,其特征在于一个标记与所述检测寡核苷酸的3'末端结合,其特征还在于另一个标记在内部结合 或在所述检测寡核苷酸的5'末端,并监测所述荧光报告的荧光 至少在多个放大循环之后。
    • 5. 发明申请
    • Detection format for hot start real time polymerase chain reaction
    • 检测格式为热启动实时聚合酶链反应
    • US20050037410A1
    • 2005-02-17
    • US10903992
    • 2004-07-30
    • Dieter HeindlWaltraud AnkenbauerFrank Laue
    • Dieter HeindlWaltraud AnkenbauerFrank Laue
    • G01N33/58C12N15/09C12P19/34C12Q1/68
    • C12Q1/6818C12Q2561/113C12Q2531/113C12Q2521/319
    • The present invention is directed to a method and a composition for amplifying and detecting a target nucleic comprising subjecting said target nucleic acid to a real time PCR amplification reaction in the presence of a thermostable DNA polymerase, a thermostable double strand dependent 3′-5′ exonuclease having a temperature optimum above 37° C., a pair of amplification primers, deoxynucleoside triphosphates, a detecting oligonucleotide carrying a first label and a second label, said first label being capable of acting as a fluorescent reporter entity when excited with light of an appropriate wavelength, said second label being capable of acting as a fluorescence quenching entity of said fluorescent reporter entity, characterized in that one label is bound to the 3′ end of said detecting oligonucleotide, and further characterized in that the other label is bound either internally or at the 5′ end of said detecting oligonucleotide, and monitoring fluorescence of said fluorescent reporter entity at least after a plurality of amplification cycles.
    • 本发明涉及用于扩增和检测靶核酸的方法和组合物,其包括在热稳定性DNA聚合酶,耐热双链依赖性3'-5'的存在下使所述靶核酸进行实时PCR扩增反应, 温度最高在37℃以上的外切核酸酶,一对扩增引物,脱氧核苷三磷酸,携带第一标记和第二标记的检测寡核苷酸,所述第一标记当用 所述第二标记能够用作所述荧光报道实体的荧光猝灭实体,其特征在于一个标记与所述检测寡核苷酸的3'末端结合,其特征还在于另一个标记在内部结合 或在所述检测寡核苷酸的5'末端,并监测所述荧光报告的荧光 至少在多个放大循环之后。