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    • 2. 发明公开
    • METHOD FOR DETERMINING NUCLEIC ACIDS BY REAL-TIME POLYMERASE CHAIN REACTION AND A DEVICE FOR THE IMPLEMENTATION THEREOF
    • 用于确定NUCLEIC通过实时聚合酶链反应和执行装置在其
    • EP2348123A1
    • 2011-07-27
    • EP09822256.5
    • 2009-10-08
    • Stroganov, Alexander AnatolievichSlyadnev, Maksim Nikolaevich
    • Stroganov, Alexander AnatolievichSlyadnev, Maksim Nikolaevich
    • C12P19/34C12Q1/68C12Q1/25
    • C12Q1/6844C12Q2565/113C12Q2565/629
    • The invention refers to molecular biology, medicine, biotechnology and is related to performance of Polymerase Chain Reaction and device for its implementation with real-time registration of reaction-product build-up. The task of the invention is solved as a result of using the method and device for its implementation for identification of nucleic acids by a real-time polymerase chain reaction including introduction of liquid samples containing nucleic acid into the reaction zones on the upper surface of the heat-conducting substrate of the microchip; isolation of the introduced samples from the atmosphere; contact of the nucleic acid of the sample with components of the polymerase chain reaction during thermocycling of the samples with heat removal through the outer surface of the microchip; fluorescent detection of the change of the quantity of the polymerase chain reaction products during thermocycling; identification of the quantity of the initial nucleic acid in the samples by the dynamic of growth of the fluorescent signal wherein the microchip used contains a heat-conducting substrate from a heat-conducting material with the thermal conductivity coefficient of more than 1 W/cm·K and the thermal diffusivity coefficient of more than 0.6 cm 2 /s while the reaction zones on the microchip surface are separated from the heat-conducting substrate by a layer of the passivating material covalently bound to the surface of the heat-conducting substrate, and the introduced samples are isolated by separating them from the atmosphere by a layer of liquid immiscible with water that is retained on the upper surface of the heat-conducting substrate by means of a peripheral barrier, with the ratio of the aggregate thermal mass of the microchip with the introduced samples and the layer of the liquid immiscible with water to thermal conductance of the microchip substrate not exceeding 0.04 s. The technical result is reduced duration of the analysis, higher reliability, accuracy, efficiency and cost-effectiveness of the analysis. 27 claims, 5 Figs.
    • 本发明涉及用于分子生物学,医学,生物技术和涉及聚合酶链反应和设备,用于与反应副产物积聚的实时登记执行性能。 本发明的任务是解决如使用其执行用于核酸的识别的方法和装置通过实时聚合酶链反应包含导入的含有核酸导入反应区上的上表面的液体样品的结果 导热的微芯片的衬底; 从大气中引入的样品的隔离; 通过微芯片的外表面热移除样品的热循环过程中与聚合酶链反应的成分的试样的核酸的接触; 聚合酶链反应产物的热循环过程中的量的变化的荧光检测; 初始核酸通过worin所使用的微芯片的荧光信号的生长的动力学样品中的量的识别包含从导热材料导热衬底具有大于1瓦/平方厘米的热导率系数· K和而在微芯片表面上的反应区从所述导热基片通过启动共价结合至所述导热基片的表面材料的负债的层分隔开的超过0.6平方厘米/秒的热扩散系数,并 所述引入的样品通过由外围挡板的装置通过与水液体不混溶的并保留在导热衬底的上表面上的层从大气中分离它们,与所述微芯片的总热质量的比率分离 与引入的样品和不超过0:04 S上的微芯片基板的热传导的液体不与水混合的层。 技术结果是减少了分析的持续时间,较高的可靠性,精确度,效率和分析的成本效益。 27和权利要求书,图5。
    • 3. 发明授权
    • METHOD FOR DETERMINING NUCLEIC ACIDS BY REAL-TIME POLYMERASE CHAIN REACTION AND A DEVICE FOR THE IMPLEMENTATION THEREOF
    • 用于确定NUCLEIC通过实时聚合酶链反应和执行装置在其
    • EP2348123B1
    • 2017-02-01
    • EP09822256.5
    • 2009-10-08
    • Stroganov, Alexander AnatolievichSlyadnev, Maksim Nikolaevich
    • Stroganov, Alexander AnatolievichSlyadnev, Maksim Nikolaevich
    • C12P19/34C12Q1/68C12Q1/25
    • C12Q1/6844C12Q2565/113C12Q2565/629
    • The invention refers to molecular biology, medicine, biotechnology and is related to performance of Polymerase Chain Reaction and device for its implementation with real-time registration of reaction-product build-up. The task of the invention is solved as a result of using the method and device for its implementation for identification of nucleic acids by a real-time polymerase chain reaction including introduction of liquid samples containing nucleic acid into the reaction zones on the upper surface of the heat-conducting substrate of the microchip; isolation of the introduced samples from the atmosphere; contact of the nucleic acid of the sample with components of the polymerase chain reaction during thermocycling of the samples with heat removal through the outer surface of the microchip; fluorescent detection of the change of the quantity of the polymerase chain reaction products during thermocycling; identification of the quantity of the initial nucleic acid in the samples by the dynamic of growth of the fluorescent signal wherein the microchip used contains a heat-conducting substrate from a heat-conducting material with the thermal conductivity coefficient of more than 1 W/cm·K and the thermal diffusivity coefficient of more than 0.6 cm 2 /s while the reaction zones on the microchip surface are separated from the heat-conducting substrate by a layer of the passivating material covalently bound to the surface of the heat-conducting substrate, and the introduced samples are isolated by separating them from the atmosphere by a layer of liquid immiscible with water that is retained on the upper surface of the heat-conducting substrate by means of a peripheral barrier, with the ratio of the aggregate thermal mass of the microchip with the introduced samples and the layer of the liquid immiscible with water to thermal conductance of the microchip substrate not exceeding 0.04 s. The technical result is reduced duration of the analysis, higher reliability, accuracy, efficiency and cost-effectiveness of the analysis. 27 claims, 5 Figs.