WO2012055073A1 - Method for setting temperature of polymerase chain reaction and instrument thereof - Google Patents

Method for setting temperature of polymerase chain reaction and instrument thereof Download PDF

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Publication number
WO2012055073A1
WO2012055073A1 PCT/CN2010/001730 CN2010001730W WO2012055073A1 WO 2012055073 A1 WO2012055073 A1 WO 2012055073A1 CN 2010001730 W CN2010001730 W CN 2010001730W WO 2012055073 A1 WO2012055073 A1 WO 2012055073A1
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Prior art keywords
temperature
polymerase chain
chain reaction
heater
reaction according
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PCT/CN2010/001730
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French (fr)
Chinese (zh)
Inventor
苏城
邓秉华
陈柏桦
李馥君
吴思颖
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瑞基海洋生物科技股份有限公司
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Priority to PCT/CN2010/001730 priority Critical patent/WO2012055073A1/en
Priority to CN201080069901.6A priority patent/CN103282496B/en
Publication of WO2012055073A1 publication Critical patent/WO2012055073A1/en

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/26—Preparation of nitrogen-containing carbohydrates
    • C12P19/28—N-glycosides
    • C12P19/30—Nucleotides
    • C12P19/34—Polynucleotides, e.g. nucleic acids, oligoribonucleotides

Definitions

  • the present invention relates to polymerase chain reaction (PCR), and more particularly to a method and apparatus for temperature setting of a polymerase chain reaction. current technology
  • PCR Polymerase chain reaction
  • This technique generally consists of 20 to 35 thermal cycles, each cycle consisting of the following three different temperature conditions: DNA denaturation at 95 ° C (Denaturation;), primer adhesion at 55 ° C ( Annealing), and an extension reaction was carried out at 72 °C.
  • the general polymerase chain reaction (PCR) is carried out in a thermal cycle apparatus that adjusts the temperature of a medium (including gas, liquid or solid;) to change the temperature of the entire mixture in the capillary tube. Polymerase chain reaction.
  • a primary object of the present invention is to provide a temperature setting method for a polymerase chain reaction which shortens the time required for a polymerase chain reaction.
  • Another object of the present invention is to provide a temperature setting device for a polymerase chain reaction which is low in cost and has great market potential.
  • the present invention provides a temperature setting method for a polymerase chain reaction, which comprises the following steps: (a) placing a test tube containing a mixed solution and a DNA sample in a closed casing (b) Heat the mixture at the bottom of the tube to a denaturation temperature with a heater.
  • the heating mode of the step (b) is a heat conduction mode.
  • the heating mode of the step (b) is a heat radiation mode.
  • the denaturation temperature is between 90 and 98 °C.
  • the invention provides a temperature setting device for polymerase chain reaction, which is used for setting a test tube
  • the temperature of the mixed liquid is characterized in that the temperature setting means comprises: a sealed casing; a heater located in the closed casing and for heating the mixed liquid located at the bottom of the test tube.
  • the heater heats the mixture at the bottom of the tube to 90 to 98 °C.
  • the heater has at least one receptacle for receiving the bottom of the test tube, and the receptacle has an imaginary axis substantially perpendicular to a horizontal plane.
  • the pocket is recessed inwardly from the side of the heater in a semi-cylindrical shape.
  • the cuvette is recessed from the top surface of the heater to have a cylindrical shape.
  • a heat insulating layer is further disposed on the top surface of the heater, and the heat insulating layer has at least one hollow portion, and the hollow portion is located at a position corresponding to the cavity of the heater.
  • the invention uses a heater to heat the mixture at the bottom of the test tube, so that the mixed liquid in the test tube generates heat convection due to the temperature gradient, so that the mixed liquid continuously changes the temperature during the convection process, and the polymerase chain reaction can be convected with the mixture. Loop through. Thereby, the time required for the conventional method to change the temperature can be greatly shortened. Moreover, the temperature setting device using the method has a simple structure and low cost, and thus has great market potential.
  • Figure 1 is a cross-sectional view showing a first preferred embodiment of the present invention
  • Figure 2 is a partial perspective view of a first preferred embodiment of the present invention
  • Figure 3 is a partial perspective view of a second preferred embodiment of the present invention.
  • the temperature setting device 10 for polymerase chain reaction provided by the first preferred embodiment of the present invention is used for setting the temperature of the mixed liquid in a plurality of test tubes T, and setting the temperature.
  • the device 10 is comprised of a hermetic housing 20, a heater 30, and a thermal insulation layer 40.
  • the sealed casing 20 has an upper cover 21, a lower cover 23, and an accommodation space 25 defined by the upper cover 21 and the lower cover 23 for accommodating a plurality of test tubes T.
  • the sealed casing 20 can shield the outside airflow from the outside airflow and affect the temperature inside the sealed casing 20.
  • the heater 30 is disposed in the hermetic casing 20 and is made of a metal having good thermal conductivity.
  • the heater 30 has a plurality of semi-cylindrical cavities 31 recessed inwardly from the side of the heater 30 to accommodate the respective test tubes.
  • the bottom portion, and each of the pockets 31 has an imaginary axis 32 that is perpendicular or approximately perpendicular to a horizontal plane.
  • Heating The device 30 can be set to have a temperature of about 90 to 98 ° C as needed, and heat the mixture located at the bottom of the test tube T to a denaturing temperature by heat conduction.
  • the heat insulating layer 40 covers the top surface of the heater 30, and the heat insulating layer 40 has a plurality of hollow portions 41 corresponding to the respective pockets 31 of the heater 30 for the test tube T to penetrate.
  • the heat insulating layer 40 blocks the heat of the heater 30 so that the temperature of the mixture in the test tube T above the heat insulating layer 40 is not affected by the heater 30.
  • a plurality of test tubes T containing the mixture and the DNA sample can be placed on a holder C (this is a conventional technique, which is not described above), so that each tube T passes through the heat insulation.
  • the hollow portion 41 of the layer 40 has its bottom end accommodated in the cavity 31 of the heater 30, and the heater 30 can heat the mixture at the bottom of the test tube T to a denaturation temperature, usually 90 to 98 ° C, in this embodiment. In the example, it is about 95 ° C.
  • the mixture in the test tube T generates heat convection due to the temperature gradient, causing the mixture at the bottom to flow upward and gradually cool down.
  • the liquid level of the mixed liquid in the test tube ⁇ is approximately equal to room temperature (about 25 ° C), and the mixture starts to heat due to the temperature gradient as the mixture at the bottom of the test tube T is heated. Convection, the liquid level of the mixed liquid will also gradually rise.
  • the mixed liquid in the test tube T will have a bonding temperature just at the primer (for example, 55 ° C;), where the primer can be bonded, and
  • the mixture in the test tube T also has another temperature (about 72 ° C) in which the primer is subjected to the extension reaction due to the temperature gradient, where the extension reaction can be carried out while the mixture continues to convect downward to the bottom of the test tube T.
  • the denaturation reaction can be carried out again, and the cycle is repeated to achieve rapid replication of specific DNA fragments for detection purposes.
  • the liquid level of the mixture will gradually rise above 55 °C, exceeding the bonding temperature of the primer, but in fact, the mixture convection cycle before the temperature rises to 55 °C.
  • the number of times is sufficient to complete the polymerase chain reaction. Thereby, the time required for the conventional method to change the temperature can be greatly shortened, and the temperature setting device using the method has a simple structure, low manufacturing cost, and great market potential.
  • the aforementioned mixture contains a buffer solution, a base pair (; dNTPs), a polymerization ta per polymer (Taq polymerase), and a primer.
  • the structure of the heater in the foregoing embodiment can be varied.
  • a temperature setting device according to a second preferred embodiment of the present invention, wherein the receptacle 51 of the heater 50 is recessed from the top surface of the heater 50 to have a cylindrical shape.
  • the heating method of the heater may be changed to heat the mixed liquid by heat radiation as needed, for example, by microwave heating.
  • the number of tanks of the heater can be set one or one as needed.
  • the number of the hollow portions of the heat insulating layer corresponding thereto is also the same.

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Abstract

A method for setting temperature of polymerase chain reaction is provided, comprising placing a tube with mixed liquor and DNA sample in an airtight shell, and heating the mixed liquor located in the bottom of the tube to denaturation temperature by using a heater. Thermal convection may take place in mixed liquor due to temperature gradient, and make the temperature of mixed liquor alter continuously during the convection process, thus polymerase chain reaction may take place circularly along with the convection of mixed liquor. Using the method of the present invention, time of polymerase chain reaction can be shortened, and cost of temperature setting instrument is low.

Description

聚合酶连锁反应的温度设定方法及装置 技术领域  Temperature setting method and device for polymerase chain reaction
本发明与聚合酶连锁反应 (PCR)有关，特别是有关于一种聚合酶连锁反应的 温度设定方法及装置。 现有技术  The present invention relates to polymerase chain reaction (PCR), and more particularly to a method and apparatus for temperature setting of a polymerase chain reaction. current technology
聚合酶连锁反应 (PCR)用于扩增特定 DNA片段， 被广泛地应用在医学及生 物学的实验室。 此项技术一般由 20〜35个热循环组成， 每个循环包含以下三个 不同温度条件的步骤： 于 95°C下进行 DNA变性反应 (Denaturation；)、 于 55°C下 进行引子黏合反应 (Annealing)、 以及于 72°C下进行延展反应 (Extension)。一般聚 合酶连锁反应 (PCR)是在一热循环设备中进行， 该热循环设备通过调整一介质 (；包括气体、液体或固体;)的温度来改变毛细试管中整体混合液的温度， 来进行聚 合酶连锁反应。  Polymerase chain reaction (PCR) is used to amplify specific DNA fragments and is widely used in medical and biological laboratories. This technique generally consists of 20 to 35 thermal cycles, each cycle consisting of the following three different temperature conditions: DNA denaturation at 95 ° C (Denaturation;), primer adhesion at 55 ° C ( Annealing), and an extension reaction was carried out at 72 °C. The general polymerase chain reaction (PCR) is carried out in a thermal cycle apparatus that adjusts the temperature of a medium (including gas, liquid or solid;) to change the temperature of the entire mixture in the capillary tube. Polymerase chain reaction.
然而， 传统的热循环设备在各步骤间的温度转换都需要相当时间， 才能改 变介质温度并使混合液与介质达到热平衡， 导致反应过程耗时冗长； 再者， 由 于传统的热循环设备必须用温控装置改变介质的温度， 导致热循环设备的价格 居高不下。 发明内容  However, the traditional thermal cycle equipment requires a considerable amount of time to change the temperature between the steps in order to change the temperature of the medium and achieve a thermal equilibrium between the mixture and the medium, which leads to a long and cumbersome reaction process. Furthermore, since the conventional thermal cycle equipment must be used The temperature control device changes the temperature of the medium, resulting in a high price of the thermal cycle equipment. Summary of the invention
本发明的主要目的是提供一种聚合酶连锁反应的温度设定方法， 其可缩短 聚合酶连锁反应所需的时间。  SUMMARY OF THE INVENTION A primary object of the present invention is to provide a temperature setting method for a polymerase chain reaction which shortens the time required for a polymerase chain reaction.
本发明的另一目的在于提供一种聚合酶连锁反应的温度设定装置， 其成本 低廉， 极具市场潜力。  Another object of the present invention is to provide a temperature setting device for a polymerase chain reaction which is low in cost and has great market potential.
为达到上述目的， 本发明所提供的一种聚合酶连锁反应的温度设定方法， 其特征在于包含有以下步骤： （a) 将一装有混合液与 DNA样品的试管置于一密 闭壳体内； （b) 用一加热器将位于所述试管底部的混合液加热至变性温度。  In order to achieve the above object, the present invention provides a temperature setting method for a polymerase chain reaction, which comprises the following steps: (a) placing a test tube containing a mixed solution and a DNA sample in a closed casing (b) Heat the mixture at the bottom of the tube to a denaturation temperature with a heater.
所述步骤 (b)的加热方式为热传导方式。  The heating mode of the step (b) is a heat conduction mode.
所述步骤 (b)的加热方式为热辐射方式。  The heating mode of the step (b) is a heat radiation mode.
所述变性温度在 90〜98°C。  The denaturation temperature is between 90 and 98 °C.
本发明所提供的一种聚合酶连锁反应的温度设定装置， 用于设定一试管中 混合液的温度， 其特征在于所述温度设定装置包含有： 一密闭壳体； 一加热器， 位于所述密闭壳体内且用于加热位于所述试管底部的混合液。 The invention provides a temperature setting device for polymerase chain reaction, which is used for setting a test tube The temperature of the mixed liquid is characterized in that the temperature setting means comprises: a sealed casing; a heater located in the closed casing and for heating the mixed liquid located at the bottom of the test tube.
所述加热器将位于所述试管底部的混合液加热至 90〜98°C。  The heater heats the mixture at the bottom of the tube to 90 to 98 °C.
所述加热器具有至少一容槽供容置所述试管底部， 且所述容槽具有一假想 轴线实质上垂直于一水平面。  The heater has at least one receptacle for receiving the bottom of the test tube, and the receptacle has an imaginary axis substantially perpendicular to a horizontal plane.
所述容槽由所述加热器的侧面向内凹设呈半圆柱状。  The pocket is recessed inwardly from the side of the heater in a semi-cylindrical shape.
所述容槽由所述加热器的顶面向下凹设而呈圆柱状。  The cuvette is recessed from the top surface of the heater to have a cylindrical shape.
还包含一绝热层覆盖于所述加热器的顶面，所述绝热层具有至少一镂空部， 所述镂空部的位置对应于所述加热器的容槽。  A heat insulating layer is further disposed on the top surface of the heater, and the heat insulating layer has at least one hollow portion, and the hollow portion is located at a position corresponding to the cavity of the heater.
本发明利用加热器对试管底部的混合液加热， 使试管中的混合液因温度梯 度而产生热对流， 使混合液在对流过程中不断改变温度， 聚合酶连锁反应即可 随着混合液对流而循环进行。 由此， 可大幅缩短传统方法变换温度所需的时间。 而且， 应用此方法的温度设定装置结构简单， 成本低廉， 因此极具市场潜力。 附图说明  The invention uses a heater to heat the mixture at the bottom of the test tube, so that the mixed liquid in the test tube generates heat convection due to the temperature gradient, so that the mixed liquid continuously changes the temperature during the convection process, and the polymerase chain reaction can be convected with the mixture. Loop through. Thereby, the time required for the conventional method to change the temperature can be greatly shortened. Moreover, the temperature setting device using the method has a simple structure and low cost, and thus has great market potential. DRAWINGS
图 1 是本发明第一较佳实施例的剖视图  Figure 1 is a cross-sectional view showing a first preferred embodiment of the present invention
图 2 是本发明第一较佳实施例的局部立体图  Figure 2 is a partial perspective view of a first preferred embodiment of the present invention
图 3 是本发明第二较佳实施例的局部立体图 本发明最佳实施方式  Figure 3 is a partial perspective view of a second preferred embodiment of the present invention.
为了详细说明本发明的结构、 特点及功效， 现举以下二较佳实施例并配合 附图说明如下。  In order to explain the structure, features and effects of the present invention in detail, the following preferred embodiments will be described below with reference to the accompanying drawings.
如图 1及图 2所示， 为本发明第一较佳实施例所提供的聚合酶连锁反应的 温度设定装置 10， 其用于设定复数个试管 T中混合液的温度， 温度设定装置 10 由一密闭壳体 20、 一加热器 30以及一绝热层 40组成。  As shown in FIG. 1 and FIG. 2, the temperature setting device 10 for polymerase chain reaction provided by the first preferred embodiment of the present invention is used for setting the temperature of the mixed liquid in a plurality of test tubes T, and setting the temperature. The device 10 is comprised of a hermetic housing 20, a heater 30, and a thermal insulation layer 40.
密闭壳体 20具有一上盖 21、 一下盖 23以及一由上盖 21与下盖 23共同界 定出的容置空间 25， 用于容置多个试管 T。 密闭壳体 20可隔绝外界气流， 避免 外界气流影响密闭壳体 20内的温度。  The sealed casing 20 has an upper cover 21, a lower cover 23, and an accommodation space 25 defined by the upper cover 21 and the lower cover 23 for accommodating a plurality of test tubes T. The sealed casing 20 can shield the outside airflow from the outside airflow and affect the temperature inside the sealed casing 20.
加热器 30设于密闭壳体 20中， 由具有良好热传导性的金属制成， 加热器 30具有复数个由加热器 30的侧面向内凹设的半圆柱状容槽 31， 以容置各个试 管 Τ的底部， 且各容槽 31具有一假想轴线 32垂直或约垂直于一水平面。 加热 器 30可依需要设定其温度大约为 90〜98°C， 并通过热传导加热位于试管 T底 部的混合液至变性温度 (denaturing temperature)。 The heater 30 is disposed in the hermetic casing 20 and is made of a metal having good thermal conductivity. The heater 30 has a plurality of semi-cylindrical cavities 31 recessed inwardly from the side of the heater 30 to accommodate the respective test tubes. The bottom portion, and each of the pockets 31 has an imaginary axis 32 that is perpendicular or approximately perpendicular to a horizontal plane. Heating The device 30 can be set to have a temperature of about 90 to 98 ° C as needed, and heat the mixture located at the bottom of the test tube T to a denaturing temperature by heat conduction.
绝热层 40覆盖在加热器 30的顶面， 绝热层 40具有复数个镂空部 41， 其 位置分别对应于加热器 30的各个容槽 31， 以供试管 T穿入。 绝热层 40可阻绝 加热器 30的热能， 使位于绝热层 40上方的试管 T内的混合液温度不致受加热 器 30影响。  The heat insulating layer 40 covers the top surface of the heater 30, and the heat insulating layer 40 has a plurality of hollow portions 41 corresponding to the respective pockets 31 of the heater 30 for the test tube T to penetrate. The heat insulating layer 40 blocks the heat of the heater 30 so that the temperature of the mixture in the test tube T above the heat insulating layer 40 is not affected by the heater 30.
实际操作该装置时， 可将复数个装有混合液 (； mixture)与 DNA样品的试管 T 放置于一固定架 C (此为习知技术， 容不赘述)上， 使各个试管 T穿过绝热层 40 的镂空部 41而使其底端容置于加热器 30的容槽 31中， 加热器 30可将试管 T 底部的混合液加热至变性温度， 通常为 90〜98°C， 在本实施例中为约 95°C， 如 此， 试管 T中混合液会因温度梯度而产生热对流， 使位于底部的混合液向上流 动并逐渐降温。 详而言之， 一开始加热时， 试管 τ中混和液的液面温度约等同 于室温 （约 25 °C )， 随着试管 T底部的混合液加热， 使混合液开始因温度梯度 而产生热对流， 混合液的液面温度也会逐渐上升， 此时， 试管 T中的混合液会 有一段恰处于引子的黏合温度（如 55°C；)， 在此即可进行引子的黏合， 并且， 试 管 T中的混合液也会有另一段因温度梯度而处于引子进行延展反应的温度 （约 72°C )， 在此即可进行延展反应， 而当混合液继续向下对流至试管 T底部时， 温 度到达 95°C，即可再次进行变性反应，如此不断循环，达到快速地复制特定 DNA 片段以供检测用的目的。 当然， 随着不断的热循环， 混合液的液面温度会逐渐 升高至 55°C以上，超过引子的黏合温度，但事实上，早在温度升高至 55°C以前， 混合液对流循环次数已足够使聚合酶连锁反应完成。 由此， 可大幅缩短传统方 法变换温度所需的时间， 且应用此方法的温度设定装置结构简单， 制造成本低 廉， 极具市场潜力。  When the device is actually operated, a plurality of test tubes T containing the mixture and the DNA sample can be placed on a holder C (this is a conventional technique, which is not described above), so that each tube T passes through the heat insulation. The hollow portion 41 of the layer 40 has its bottom end accommodated in the cavity 31 of the heater 30, and the heater 30 can heat the mixture at the bottom of the test tube T to a denaturation temperature, usually 90 to 98 ° C, in this embodiment. In the example, it is about 95 ° C. Thus, the mixture in the test tube T generates heat convection due to the temperature gradient, causing the mixture at the bottom to flow upward and gradually cool down. In detail, at the beginning of heating, the liquid level of the mixed liquid in the test tube τ is approximately equal to room temperature (about 25 ° C), and the mixture starts to heat due to the temperature gradient as the mixture at the bottom of the test tube T is heated. Convection, the liquid level of the mixed liquid will also gradually rise. At this time, the mixed liquid in the test tube T will have a bonding temperature just at the primer (for example, 55 ° C;), where the primer can be bonded, and The mixture in the test tube T also has another temperature (about 72 ° C) in which the primer is subjected to the extension reaction due to the temperature gradient, where the extension reaction can be carried out while the mixture continues to convect downward to the bottom of the test tube T. When the temperature reaches 95 °C, the denaturation reaction can be carried out again, and the cycle is repeated to achieve rapid replication of specific DNA fragments for detection purposes. Of course, with continuous thermal cycling, the liquid level of the mixture will gradually rise above 55 °C, exceeding the bonding temperature of the primer, but in fact, the mixture convection cycle before the temperature rises to 55 °C. The number of times is sufficient to complete the polymerase chain reaction. Thereby, the time required for the conventional method to change the temperature can be greatly shortened, and the temperature setting device using the method has a simple structure, low manufacturing cost, and great market potential.
在本发明中， 前述的混合液包含有缓冲溶液 (； buffer)、碱基对 (； dNTPs；)、 聚合 酉每 (Taq polymerase)及弓 |子 (primers)。  In the present invention, the aforementioned mixture contains a buffer solution, a base pair (; dNTPs), a polymerization ta per polymer (Taq polymerase), and a primer.
依据本发明的精神， 前述实施例中加热器的结构可加以变化。如图 3所示， 为本发明第二较佳实施例所提供的温度设定装置， 其中， 加热器 50 的容槽 51 由加热器 50的顶面向下凹设而呈圆柱状。 事实上， 加热器的加热方式除了前述 的热传导方式之外， 也可根据需要变化为以热辐射的方式对混合液进行加热， 例如通过微波方式进行加热。  In accordance with the spirit of the present invention, the structure of the heater in the foregoing embodiment can be varied. As shown in Fig. 3, a temperature setting device according to a second preferred embodiment of the present invention, wherein the receptacle 51 of the heater 50 is recessed from the top surface of the heater 50 to have a cylindrical shape. In fact, in addition to the aforementioned heat transfer mode, the heating method of the heater may be changed to heat the mixed liquid by heat radiation as needed, for example, by microwave heating.
在此， 需补充说明的是， 加热器的容槽的数目， 可根据需要设一个或一个 以上， 与其对应的绝热层的镂空部的数目也相同。 Here, it should be added that the number of tanks of the heater can be set one or one as needed. The number of the hollow portions of the heat insulating layer corresponding thereto is also the same.
本发明在前述实施例中所揭示的构成组件， 仅为举例说明， 并不能用来限 制本案的专利范围， 其它等效组件的替代或变化， 均应被涵盖在本案的专利保 护范围内。  The components of the present invention disclosed in the foregoing embodiments are merely illustrative and are not intended to limit the scope of the patents of the present invention. Other alternatives or variations of equivalent components are to be covered by the patent protection of the present invention.

Claims

权利要求 Rights request
1、 一种聚合酶连锁反应的温度设定方法， 其特征在于包含有以下步骤：A method for setting a temperature of a polymerase chain reaction, which comprises the steps of:
(a) 将一装有混合液与 DNA样品的试管置于一密闭壳体内； (a) placing a test tube containing the mixture and DNA sample in a closed housing;
(b) 用一加热器将位于所述试管底部的混合液加热至变性温度。  (b) Heating the mixture at the bottom of the tube to a denaturation temperature with a heater.
2、 如权利要求 1所述聚合酶连锁反应的温度设定方法， 其特征在于： 所述 步骤 (b)的加热方式为热传导方式。  The temperature setting method of the polymerase chain reaction according to claim 1, wherein the heating method of the step (b) is a heat conduction method.
3、 如权利要求 1所述聚合酶连锁反应的温度设定方法， 其特征在于： 所述 步骤 (b)的加热方式为热辐射方式。  The method for setting a temperature of a polymerase chain reaction according to claim 1, wherein the heating method of the step (b) is a heat radiation method.
4、 如权利要求 1所述聚合酶连锁反应的温度设定方法， 其特征在于： 所述 变性温度在 90〜98°C。  A method for setting a temperature of a polymerase chain reaction according to claim 1, wherein the denaturation temperature is 90 to 98 °C.
5、一种聚合酶连锁反应的温度设定装置，用于设定一试管中混合液的温度， 其特征在于所述温度设定装置包含有：  5. A temperature setting device for a polymerase chain reaction for setting a temperature of a mixed solution in a test tube, characterized in that said temperature setting means comprises:
一密闭壳体；  a closed casing;
一加热器， 位于所述密闭壳体内且用于加热位于所述试管底部的混合液。 A heater is located within the containment housing and is used to heat the mixture located at the bottom of the tube.
6、 如权利要求 5所述聚合酶连锁反应的温度设定装置， 其特征在于： 所述 加热器将位于所述试管底部的混合液加热至 90〜98°C。 A temperature setting device for a polymerase chain reaction according to claim 5, wherein: said heater heats the mixture at the bottom of said test tube to 90 to 98 °C.
7、 如权利要求 5所述聚合酶连锁反应的温度设定装置， 其特征在于： 所述 加热器具有至少一容槽供容置所述试管底部， 且所述容槽具有一假想轴线实质 上垂直于一水平面。  7. The temperature setting device for a polymerase chain reaction according to claim 5, wherein: said heater has at least one cuvette for accommodating said test tube bottom, and said cuvette has an imaginary axis substantially Vertical to a horizontal plane.
8、 如权利要求 7所述聚合酶连锁反应的温度设定装置， 其特征在于： 所述 容槽由所述加热器的侧面向内凹设呈半圆柱状。  8. The temperature setting device for a polymerase chain reaction according to claim 7, wherein: said pocket is recessed inwardly from a side surface of said heater in a semi-cylindrical shape.
9、 如权利要求 7所述聚合酶连锁反应的温度设定装置， 其特征在于： 所述 容槽由所述加热器的顶面向下凹设而呈圆柱状。  A temperature setting device for a polymerase chain reaction according to claim 7, wherein: said pocket is recessed from a top surface of said heater to have a cylindrical shape.
10、 如权利要求 7所述聚合酶连锁反应的温度设定装置， 其特征在于： 还 包含一绝热层覆盖于所述加热器的顶面， 所述绝热层具有至少一镂空部， 所述 镂空部的位置对应于所述加热器的容槽。  10. The temperature setting device of the polymerase chain reaction according to claim 7, further comprising: a heat insulating layer covering a top surface of the heater, the heat insulating layer having at least one hollow portion, the hollowing out The position of the portion corresponds to the pocket of the heater.
PCT/CN2010/001730 2010-10-29 2010-10-29 Method for setting temperature of polymerase chain reaction and instrument thereof WO2012055073A1 (en)

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