JP5910304B2 - Heat cycle equipment - Google Patents

Description

  The present invention relates to a heat cycle apparatus.

  In recent years, gene-based medical care such as gene diagnosis and gene therapy has attracted attention due to the development of gene utilization technology, and many methods using genes for variety discrimination and variety improvement have been developed in the field of agriculture and livestock. . Techniques such as PCR (Polymerase Chain Reaction) are widely used as techniques for utilizing genes. Today, PCR has become an indispensable technique for elucidating information on biological materials.

  The PCR method is a technique for amplifying a target nucleic acid by subjecting a solution (reaction solution) containing a nucleic acid (target nucleic acid) to be amplified and a reagent to thermal cycling. The thermal cycle is a process in which two or more stages of temperature are periodically applied to the reaction solution. In the PCR method, a method of performing a two-stage or three-stage thermal cycle is common.

  In the PCR method, a container for performing a biochemical reaction, generally called a tube or a biological sample reaction chip (biochip), is used. However, the conventional methods have a problem that the amount of reagents and the like necessary for the reaction is large, the apparatus is complicated to realize a thermal cycle necessary for the reaction, and the reaction takes time. Therefore, a biochip and a reaction apparatus are required for performing PCR accurately and in a short time using a very small amount of reagent or specimen.

  In order to solve such a problem, Patent Document 1 is filled with a reaction liquid and a liquid that is immiscible with the reaction liquid and has a specific gravity smaller than that of the reaction liquid (mineral oil or the like, hereinafter referred to as “liquid”). A biological sample reaction apparatus is disclosed in which a reaction liquid is moved to perform a thermal cycle by rotating the biological sample reaction container around a horizontal rotation axis.

JP 2009-136250 A

  However, in Patent Document 1, it is preferable to reduce the thickness of the biological sample reaction container in order to shorten the time required for the reaction. On the other hand, when the container is configured to be sealed with a stopper, it is necessary to provide a stopper with a large wall thickness and sufficient size in order to improve durability against stress due to fitting and ease of fitting operation. It was. As a result, the heat capacity of the stopper becomes relatively large, the heat transfer to the liquid on the side near the stopper is slower than that on the side far from the stopper, and it takes time for the oil near the stopper to reach a predetermined temperature. It became.

  Therefore, a heat cycle device that can heat the reaction vessel to a desired temperature in a shorter time has been required.

  SUMMARY An advantage of some aspects of the invention is to solve at least a part of the problems described above, and the invention can be implemented as the following forms or application examples.

  [Application Example 1] In the heat cycle device according to this application example, the reaction liquid and the reaction liquid are filled with a liquid having a different specific gravity and immiscible with the reaction liquid, and the inner walls facing the reaction liquid A flow path that moves along the flow path; a mounting portion that mounts a reaction container that includes a sealing portion that seals the flow path; and the flow path of the flow path when the reaction container is mounted to the mounting section. A first heating section for heating the first region; a second heating section for heating the sealing section when the reaction vessel is mounted on the mounting section; the first and second heating sections; and the mounting A drive mechanism that switches the arrangement of the portion between the first arrangement and the second arrangement, and the first arrangement is arranged when the reaction container is attached to the attachment portion. 1 region is an arrangement located at the lowermost part of the flow path in the direction in which gravity acts, and the second region When the reaction vessel is mounted on the mounting portion, the second region of the flow path in which the position in the direction in which the reaction solution moves is different from the first region in the direction in which the gravity acts is It is arrangement | positioning located in the lowest part of a flow path, It is characterized by the above-mentioned.

  According to this application example, the state in which the reaction container is held in the first arrangement and the state in which the reaction container is held in the second arrangement by switching the arrangement of the first and second heating units and the mounting unit. And can be switched. The first arrangement is an arrangement in which the first region of the flow path constituting the reaction vessel is located at the lowest part of the flow path in the direction in which gravity acts. The second arrangement is an arrangement in which the second region of the flow path constituting the reaction vessel is located at the lowermost part of the flow path in the direction in which gravity acts. That is, for example, when the specific gravity of the reaction liquid is relatively large, the reaction liquid can be held in the first region in the first arrangement and the reaction liquid can be held in the second area in the second arrangement by the action of gravity. Since the first region is heated by the first heating unit, the first region and the second region can be set to different temperatures. Therefore, since the reaction liquid can be maintained at a predetermined temperature while the reaction vessel is held in the first arrangement or the second arrangement, it is possible to provide a thermal cycle apparatus capable of easily controlling the heating time. Further, since the second heating unit heats the sealed portion of the reaction vessel, the region of the reaction vessel that is relatively closer to the sealed portion is shorter than when the thermal cycle device does not include the second heating unit. A thermal cycle apparatus that can be heated to a desired temperature over time can be realized.

  Application Example 2 In the heat cycle apparatus according to the application example described above, the second heating unit is inserted into a recess formed in the sealing unit when the reaction container is mounted on the mounting unit. A protrusion may be further included.

  According to this application example, the second heating unit can heat the liquid immiscible with the reaction liquid at a closer position through the protrusion inserted into the recess provided in the sealing unit, so that the liquid can be heated in a shorter time. As a result, the reaction solution can be quickly heated in the second arrangement.

Sectional drawing of the reaction container and heating part which concern on this embodiment. The perspective view showing the state where the lid of the heat cycle device concerning this embodiment was closed. The perspective view showing the state which opened the lid | cover of the heat cycle apparatus which concerns on this embodiment. The disassembled perspective view of the main body of the heat cycle apparatus which concerns on this embodiment. (A) is sectional drawing which shows typically the cross section in the surface perpendicular to a rotating shaft through the AA line of FIG. 2 in 1st arrangement | positioning, (B) is A of FIG. 2 in 2nd arrangement | positioning. Sectional drawing which shows typically the cross section in the surface which passes along A line and is perpendicular | vertical to a rotating shaft. Sectional drawing of the reaction container and heating part which concern on embodiment of a comparison. Sectional drawing of the reaction container and heating part of another structure which concern on embodiment of a comparison.

  DESCRIPTION OF EXEMPLARY EMBODIMENTS Hereinafter, embodiments of the invention will be described with reference to the drawings. Note that the drawings to be used are appropriately enlarged or reduced so that the part to be described can be recognized.

1. Reaction Container FIG. 1 is a cross-sectional view of the reaction container 1 and the heating units 1012, 1013, and 1052 according to this embodiment.
The reaction container 1 attached to the heat cycle apparatus 1000 (see FIG. 3) of the present embodiment will be described. As shown in FIG. 1, the reaction vessel 1 includes a reaction vessel body 11 and a stopper (sealing part) 12, and a second liquid (reaction) is contained in a first chamber (flow path) 110 of the reaction vessel 1. The liquid) 40 and the second liquid 40 are filled with a first liquid (liquid) 30 having a different specific gravity.

  The stopper 12 seals the first chamber 110. The second liquid 40 moves close to the opposing inner wall of the first chamber 110. Dimethyl silicone oil or paraffin oil can be used for the first liquid 30. In particular, dimethyl silicone oil is suitable for thermal cycling because it is flame retardant. Since dimethyl silicone oil has high insulation properties, it is preferable to add a fluorinated silicone resin. Thereby, since the charging of the second liquid 40 can be prevented, the second liquid 40 can be appropriately moved.

  The reaction vessel body 11 and the stopper 12 are made of transparent polypropylene that does not inhibit the PCR reaction and is resistant to heat. Polycarbonate may be used as a resin material that has heat resistance and does not inhibit the PCR reaction. However, since it has autofluorescence, it is not suitable for performing real-time PCR by fluorescence measurement. Other than the resin material, borosilicate glass having heat resistance is suitable. The thickness of the reaction vessel main body 11 is preferably as thin as possible from the viewpoint of transparency and heat capacity (shortening of heating time), but is preferably about 0.3 to 0.5 mm from the viewpoint of moldability and strength. As for the strength, by reacting with the thermal expansion of the first liquid 30 in order to prevent the dissolved gas from foaming from the first liquid 30 or the like by heating by setting the thickness to about 0.3 to 0.5 mm. The pressure in the reaction vessel 1 is increased by increasing the rigidity of the vessel 1 (Henry's law). The inner diameter of the reaction vessel 1 is 2 to 3 mm and the length is about 15 to 25 mm. The wall thickness of the stopper 12 is about 0.7 mm which is a little thicker to ensure fitting, and the length is 8 mm in operation. This is necessary. In the present embodiment, the stopper 12 is provided with a recess 12a. In the present embodiment, the tip of the reaction vessel 1 has a mirror finished inner surface (wall surface of the first chamber 110) and outer surface for fluorescence measurement.

  The second liquid 40 is a liquid that is immiscible with the first liquid 30. As the second liquid 40, for example, a mixed liquid of a reagent containing an enzyme or a drug necessary for a desired reaction and a test liquid (a liquid containing a specimen) can be used using water as a medium. When a reaction including fluorescence measurement is performed as in real-time PCR, a second liquid 40 composed of a DNA to be amplified, a DNA polymerase, a primer, and a fluorescent dye whose luminance changes in accordance with the amplification is placed in the reaction container 1. 1 to 2 μL is dropped and sealed with a stopper 12. When fluorescence measurement is not performed, the fluorescent dye may not be included. The first liquid 30 is filled in advance until it reaches the reservoir 14 so that air bubbles do not enter when the second liquid 40 is sealed, and the first liquid 30 overflows into the reservoir 14 by the stopper 12 having a sharp tip. Fit while letting it out. The overflowing first liquid 30 is held in the reservoir 14 and sealed with a snap 13 so as not to overflow to the outside. In this way, the first chamber 110 sealed with the reaction vessel main body 11 and the stopper 12 fitted together is sealed.

2. Thermal Cycle Device FIG. 2 is a perspective view showing a state where the lid 1050 of the thermal cycle device 1000 according to the present embodiment is closed, and FIG. 3 shows a state where the lid 1050 of the thermal cycle device 1000 according to the present embodiment is opened. It is a perspective view. FIG. 4 is an exploded perspective view of the main body 1010 of the heat cycle apparatus 1000 according to the present embodiment. 5A is a cross-sectional view schematically showing a cross section in a plane perpendicular to the rotation axis R through the line AA in FIG. 2 in the first arrangement, and FIG. 5B is a second arrangement. 3 is a cross-sectional view schematically showing a cross section in a plane that passes through the line AA in FIG. 2 and is perpendicular to the rotation axis R. FIG. In FIGS. 5A and 5B, the white arrow indicates the rotation direction of the main body 1010, and the arrow g indicates the direction in which gravity acts.

  The thermal cycle apparatus 1000 shown in FIGS. 2, 3, and 4 includes a mounting portion 1011 for mounting the reaction vessel 1, and the first chamber 110 of the reaction vessel 1 when the reaction vessel 1 is mounted on the mounting portion 1011. In contrast, a temperature gradient forming unit 1030 that forms a temperature gradient in the direction in which the second liquid 40 moves (in the present embodiment, the longitudinal direction of the first chamber 110), the mounting unit 1011 and the temperature gradient forming unit. And a drive mechanism 1020 that rotates 1030 on the same rotation axis R that is an axis in a direction having a horizontal component.

  In the example shown in FIGS. 2 and 3, the thermal cycle apparatus 1000 includes a main body 1010 and a drive mechanism 1020. As shown in FIG. 4, the main body 1010 includes a mounting portion 1011 and a temperature gradient forming portion 1030.

  The mounting unit 1011 has a structure for mounting the reaction container 1. In the example shown in FIGS. 3 and 4, the mounting portion 1011 of the heat cycle apparatus 1000 has a slot structure in which the reaction vessel 1 is inserted and mounted. In the example shown in FIG. 4, the mounting unit 1011 inserts the reaction container 1 into a hole penetrating a first heat block 1012b of the first heating unit 1012 and a third heat block 1013b of the third heating unit 1013, which will be described later. It has a structure. In the example shown in FIG. 3, 20 mounting portions 1011 are provided in the main body 1010.

  The temperature gradient forming unit 1030 forms a temperature gradient in the direction in which the second liquid 40 moves with respect to the first chamber 110 of the reaction vessel 1 when the reaction vessel 1 is attached to the attachment unit 1011. Here, “forming a temperature gradient” means forming a state in which the temperature changes along a predetermined direction. Therefore, “to form a temperature gradient in the direction in which the second liquid 40 moves” means to form a state in which the temperature changes along the direction in which the second liquid 40 moves. “The state in which the temperature changes along the predetermined direction” is, for example, that the temperature may be monotonously high or low along the predetermined direction, or from a change in which the temperature increases along the predetermined direction. You may change on the way from the change which becomes low, or from the change which becomes low to the change which becomes high. In the example shown in FIGS. 3 and 4, the temperature gradient forming unit 1030 includes a first heating unit 1012, a second heating unit 1052, and a third heating unit 1013. In the main body 1010 of the heat cycle apparatus 1000, the first heating unit 1012 and the third heating unit 1013 are fixed around the periphery by a flange 1016 and a fixing plate 1019.

  The first heating unit 1012 heats the first temperature region (first region) 1111 of the first chamber 110 to the first temperature when the reaction vessel 1 is mounted on the mounting unit 1011. In the example shown in FIGS. 5A and 5B, the first heating unit 1012 is disposed in the main body 1010 at a position where the first temperature region 1111 of the first chamber 110 is heated.

  In the example shown in FIGS. 4, 5 </ b> A, and 5 </ b> B, the first heating unit 1012 includes a first heater 1012 a as a mechanism for generating heat, and a member that transmits the generated heat to the reaction vessel 1. As a first heat block 1012b. In the heat cycle apparatus 1000, the first heater 1012a is a cartridge heater, and is connected to an external power source (not shown) by a conducting wire 1015.

  When the reaction vessel 1 is mounted on the mounting unit 1011, the third heating unit 1013 moves the second temperature region (second region) 1112 of the first chamber 110 to a second temperature different from the first temperature. Heat to. In the example shown in FIGS. 5A and 5B, the third heating unit 1013 is disposed in the main body 1010 at a position where the second temperature region 1112 of the reaction vessel 1 is heated. The third heating unit 1013 includes a third heater 1013a and a third heat block 1013b. The configuration of the third heating unit 1013 is the same as that of the first heating unit 1012 except that the area of the first chamber 110 to be heated and the heating temperature are different from those of the first heating unit 1012.

  Thermal cycle apparatus 1000 includes a lid 1050. In the example shown in FIGS. 3, 5 </ b> A, and 5 </ b> B, the lid 1050 is provided so as to cover the mounting portion 1011. The lid 1050 includes a second heating unit 1052 and a cover 1053, and heats the second temperature region 1112 together with the third heating unit 1013 through a protrusion 1051 molded on the bottom surface of the lid 1050. In the example shown in FIGS. 5A and 5B, the second heating unit 1052 is disposed at a position where the stopper 12 of the reaction vessel 1 is heated when the reaction vessel 1 is attached to the attachment unit 1011. Has been. The second heating unit 1052 is disposed in the main body 1010 at a position for heating the second temperature region 1112 of the first chamber 110.

  In the example shown in FIGS. 1, 3, 5 </ b> A, and 5 </ b> B, the second heating unit 1052 supplies the second heater 1052 a as a mechanism for generating heat and the generated heat to the plug 12. The second heat block 1052b as a member to be transmitted is included. In the heat cycle apparatus 1000, the second heater 1052a is a cartridge heater, and is connected to an external power source (not shown) by a conducting wire 1015.

  The second heater 1052a is made of a metal having high thermal conductivity such as aluminum, and a plurality of protrusions 1051 shaped so as to come into contact with the recess 12a of the stopper 12 when the reaction vessel 1 is attached to the attachment portion 1011 are formed on the bottom surface of the lid 1050. Molded. As a result, the first liquid 30 is heated via the plug 12. The protrusion 1051 is preferably formed integrally with the lid 1050 in terms of heat conduction. When a high-precision finish is required so that the protrusion 1051 is in close contact with the stopper 12, it may be screwed into the lid 1050 after finishing separately. However, when screwing, it is preferable to devise measures to improve the heat conduction efficiency, such as applying silicon grease to the screw portion. In the present embodiment, the tip of the projection 1051 is made slightly longer so that it comes into close contact with the end of the hollow portion of the plug 12 so that there is no gap. The lid 1050 may be closed with the stopper 12 fitted into the reaction vessel main body 11, or the stopper 1050 is fitted to the protrusion 1051 of the lid 1050 in advance, and the lid 1050 is closed with the stopper 12 heated. The stoppers 12 can be fitted together into a plurality of reaction vessel bodies 11.

  By providing the projection 1051 on the lid 1050, the contact of the recess 12a of the stopper 12 (region close to the first chamber 110 when the first chamber 110 is sealed) and its vicinity can be heated. Therefore, the first liquid 30 can be heated from a closer position.

  According to this embodiment, the second heating unit 1052 can heat the first liquid 30 that is immiscible with the second liquid 40 at a closer position through the protrusion 1051 inserted into the recess 12 a provided in the stopper 12. As a result, the first liquid 30 can be heated to an appropriate temperature in a shorter time than the case without the protrusion 1051, and as a result, the second liquid 40 can be quickly heated in the second arrangement. it can.

  Moreover, if the thickness at the end of the recess 12a is made thinner than the other positions of the plug 12, the first liquid 30 can be heated more efficiently. In the present embodiment, the second temperature region 1112 is heated to an appropriate temperature in about 10 seconds. Since the upper surface and the side surface of the lid 1050 are heated by the heat of the second heating unit 1052, the lid 1050 is covered with a cover 1053 made of a heat insulating material so as not to be burned during the heating.

  A second heating unit 1052 and a third heating unit 1013 at a temperature of about 95 ° C. necessary for PCR thermal denaturation are provided on the stopper 12 side of the reaction vessel 1, and a first heating unit 1012 at a temperature of about 60 ° C. required for an extension reaction is provided on the tip side. By setting, different temperature zones can be provided for the first liquid 30 in the first chamber 110. At this time, since the first chamber 110 is elongated and the convection is hindered by viscous resistance, a sufficient temperature difference can be obtained even if the length of the first chamber 110 is around 15 mm.

  Since the specific gravity of the second liquid 40 is heavier than that of the first liquid 30, the second liquid 40 reciprocates in different temperature zones in the first chamber 110 when the whole is rotated on the surface including the axial direction of the reaction vessel 1. PCR reaction occurs. During the amplification and after the amplification, the DNA is quantified by irradiating the second liquid 40 with excitation light from the fluorescence detector 70 from the front end side of the reaction container 1, and the fluorescence according to the amount of DNA emitted by the fluorescent substance in the second liquid 40. Is measured by the fluorescence detector 70.

  The temperatures of the first heating unit 1012, the third heating unit 1013, and the lid 1050 (second heating unit 1052) may be controlled by a temperature sensor (not shown) and a control unit described later.

  The drive mechanism 1020 is a mechanism that rotates the mounting unit 1011 and the temperature gradient forming unit 1030 on the same rotation axis R that is an axis in a direction having a horizontal component. “Direction having a horizontal component” is expressed as a vector sum of a vertical component (component parallel to the direction in which gravity acts) and a horizontal component (component perpendicular to the direction in which gravity acts) A direction having a horizontal component. In the present embodiment, the drive mechanism 1020 includes a bearing 1021, a motor and a drive shaft (not shown), and is configured by connecting the drive shaft and the flange 1016 of the main body 1010. The bearing 1021 supports the outer periphery of the flange 1016 on the bearing 1021 side as a sliding surface. When the motor of the drive mechanism 1020 is operated, the main body 1010 is rotated with the drive shaft as the rotation axis R.

  As shown in FIG. 5A, the first arrangement is such that the end of the first chamber 110 of the reaction vessel 1 that is relatively far from the lid 1050 is the lowest point in the direction in which gravity acts. It is the arrangement which becomes. That is, in the first arrangement, when the reaction vessel 1 is mounted on the mounting portion 1011, the first temperature region 1111 of the first chamber 110 is placed at the lowermost portion of the first chamber 110 in the direction in which gravity acts. It is an arrangement to be positioned. In the example shown in FIG. 5A, in the first arrangement, the second liquid 40 having a specific gravity greater than that of the first liquid 30 exists in the first temperature region 1111. Accordingly, the second liquid 40 is placed under the first temperature.

  As shown in FIG. 5B, the second arrangement is such that the end of the first chamber 110 of the reaction vessel 1 on the side relatively close to the lid 1050 is the lowest point in the direction in which gravity acts. It is the arrangement which becomes. That is, in the second arrangement, when the reaction vessel 1 is mounted on the mounting portion 1011, the second temperature region 1112 of the first chamber 110 is placed at the lowermost portion of the first chamber 110 in the direction in which gravity acts. It is an arrangement to be positioned. In the example shown in FIG. 5B, in the second arrangement, the second liquid 40 having a specific gravity greater than that of the first liquid 30 exists in the second temperature region 1112. Therefore, the second liquid 40 is placed under the second temperature.

  As described above, the driving mechanism 1020 rotates the mounting unit 1011 and the temperature gradient forming unit 1030 between the first arrangement and the second arrangement different from the first arrangement, whereby the second liquid 40 can be heat cycled.

  The thermal cycle apparatus 1000 of this embodiment is filled with a liquid (first liquid 30 in this embodiment) that is not miscible with the reaction liquid (second liquid 40 in this embodiment) and has a specific gravity different from that of the reaction liquid. It is a device that realizes a thermal cycle by reciprocating a reaction solution contained in the form of droplets in the reaction vessel 1 between a certain temperature region in the reaction vessel 1 and another temperature region having a different temperature. .

  According to this embodiment, by switching the arrangement of the first and second heating units 1012 and 1052 and the mounting unit 1011, the reaction vessel 1 is held in the first arrangement, and the reaction vessel 1 is in the second arrangement. The state held in the arrangement can be switched. The first arrangement is an arrangement in which the first temperature region 1111 of the first chamber 110 constituting the reaction vessel 1 is located at the lowermost part of the first chamber 110 in the direction in which gravity acts. The second arrangement is an arrangement in which the second temperature region 1112 of the first chamber 110 constituting the reaction vessel 1 is located at the lowermost part of the first chamber 110 in the direction in which gravity acts. That is, for example, when the specific gravity of the second liquid 40 is relatively large, the second liquid 40 is brought into the first temperature region 1111 in the first arrangement and the second liquid 40 in the second arrangement due to the action of gravity. The second liquid 40 can be held in the second temperature region 1112. Since the first temperature region 1111 is heated by the first heating unit 1012, the first temperature region 1111 and the second temperature region 1112 can be set to different temperatures. Accordingly, since the second liquid 40 can be held at a predetermined temperature while the reaction vessel 1 is held in the first arrangement or the second arrangement, it is possible to provide the thermal cycle apparatus 1000 that can easily control the heating time. In addition, since the second heating unit 1052 heats the plug 12 of the reaction vessel 1, the region of the reaction vessel 1 that is relatively close to the plug 12 as compared with the case where the thermal cycle apparatus 1000 does not include the second heating unit 1052. Can be realized in a shorter time in a heat cycle apparatus 1000 that can be heated to a desired temperature.

FIG. 6 is a cross-sectional view of the reaction vessel 1 and the third heating unit 1013 according to the comparative embodiment, and FIG. 7 is a diagram of another embodiment of the reaction vessel 1 and the heating units 1012, 1013, and 1052 according to the comparative embodiment. FIG.
The example shown in FIG. 6 has shown the case where the cover 1050 which consists of material with high heat insulation is carried out as a comparison. The lid 1050 of FIG. 6 does not include the second heater 1052a. In this case, the stopper 12 formed with the recess 12a serves as a cooling fin to prevent shrinkage during molding, and the heat of the first liquid 30 is released to the lid 1050 side. It has been confirmed that it takes about one minute to reach When PCR is performed using a thermal cycle apparatus that moves the reaction solution between the first temperature region 1111 and the second temperature region 1112 of the first chamber 110, a series of thermal cycles takes about 15 minutes. Done. Therefore, in order to shorten the reaction time, 1 minute is unacceptable as a loss time. On the other hand, in the example shown in FIG. 7, the second heating unit 1052 including a cartridge heater with a temperature sensor is incorporated in the lid 1050 to heat the plug 12. The lid 1050 is made of a metal having high thermal conductivity such as aluminum and is covered with a cover 1053 made of a heat insulating material so as not to be burned. By incorporating the second heating unit 1052, the plug 12 can be heated, so that heat dissipation from the plug 12 can be suppressed as compared with the method shown in FIG. 6. However, in order to improve the workability of the work of sealing the second liquid 40 in the reaction vessel 1, the length of the stopper 12 protruding from the snap 13 is preferably about 8 mm or more. In this case, since the heat capacity of the plug 12 is large, it is preferable to provide a convex portion that can be heated from a closer distance in order to shorten the time.

  The heat cycle apparatus 1000 may include a control unit (not shown). The control unit controls at least one of the drive mechanism 1020 and the temperature gradient forming unit 1030. The control unit may be configured to perform control described later by being realized by a dedicated circuit. The control unit functions as a computer by a CPU (Central Processing Unit) executing a control program stored in a storage device such as a ROM (Read Only Memory) or a RAM (Random Access Memory). You may be comprised so that control may be performed.

  In addition, embodiment mentioned above and a modification are examples, Comprising: It is not necessarily limited to these. For example, a plurality of embodiments and modifications can be combined as appropriate.

  The present invention is not limited to the above-described embodiments, and various modifications can be made. For example, the present invention includes substantially the same configuration (for example, a configuration having the same function, method and result, or a configuration having the same purpose and effect) as the configuration described in the embodiment. In addition, the invention includes a configuration in which a non-essential part of the configuration described in the embodiment is replaced. In addition, the present invention includes a configuration that exhibits the same operational effects as the configuration described in the embodiment or a configuration that can achieve the same object. In addition, the invention includes a configuration in which a known technique is added to the configuration described in the embodiment.

  DESCRIPTION OF SYMBOLS 1 ... Reaction container 11 ... Reaction container main body 12 ... Plug (sealing part) 12a ... Recess 13 ... Snap 14 ... Reservoir 30 ... 1st liquid (liquid) 40 ... 2nd liquid (reaction liquid) 70 ... Fluorescence detector DESCRIPTION OF SYMBOLS 110 ... 1st chamber (flow path) 1000 ... Thermal cycle apparatus 1010 ... Main body 1011 ... Mounting part 1012 ... 1st heating part 1012a ... 1st heater 1012b ... 1st heat block 1013 ... 3rd heating part 1013a ... 3rd heater 1013b ... 3rd heat block 1015 ... Conductor 1016 ... Flange 1019 ... Fixed plate 1020 ... Drive mechanism 1021 ... Bearing 1030 ... Temperature gradient formation part 1050 ... Lid 1051 ... Projection 1052 ... 2nd heating part 1052a ... 2nd heater 1052b ... 2nd Heat block 1053 ... cover 1111 ... first temperature range (First region) 1112 ... second temperature region (second region).

Claims (4)

A reaction liquid and a liquid having a specific gravity different from that of the reaction liquid and immiscible with the reaction liquid, a container having an opening at one end and a flow path for the reaction liquid to move; and the opening and possible mounting portion mounting a reaction vessel containing a sealing portion, a having a first recess which is recessed toward the interior of the container when sealed,
A first heating unit that heats the first region of the flow path to a first temperature when the reaction vessel is mounted on the mounting unit;
A second heating unit that heats the sealing unit to a second temperature different from the first temperature when the reaction vessel is mounted to the mounting unit;
A drive mechanism for switching the arrangement of the first and second heating units and the mounting unit between the first arrangement and the second arrangement;
Including
The first arrangement is an arrangement in which, when the reaction vessel is attached to the attachment part, the first region is located at the lowest part of the flow path in the direction in which gravity acts,
In the second arrangement, when the reaction vessel is mounted on the mounting portion, the second region of the flow path in which the position in the direction in which the reaction solution moves differs from the first region is the action of gravity. Is located at the bottom of the flow path in the direction to
The container has an inner diameter changing portion whose inner diameter is reduced from the one end toward the other end of the container, and a second recess formed on the inner wall of the container and engaged with the sealing portion.
The sealing portion has an outer diameter changing portion whose outer diameter is reduced from one end of the sealing portion toward the other end, a convex portion that is formed on the outer wall of the sealing portion and engages with the container, Have
When the flow path is sealed by the sealing portion, the second concave portion and the convex portion are engaged with each other, the inner diameter changing portion and the outer diameter changing portion are fitted, and the inside of the container is accommodated. It is formed to be divided into a chamber and a reservoir,
The second heating unit is a thermal cycle apparatus including a protrusion that contacts the first recess when the reaction container is mounted on the mounting unit. The thermal cycle apparatus according to claim 1, wherein
When the reaction container is mounted on the mounting portion, the protrusion is a thermal cycle device that is screwed into the first recess. The thermal cycle apparatus according to claim 2, wherein
A thermal cycle device in which grease is applied to the protrusion. In the heat cycle apparatus according to any one of claims 1 to 3,
The thermal cycle apparatus which has a cover which covers the said 2nd heating part.

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