JP3790919B2 - Trace liquid reactor - Google Patents

Description

[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a reaction device, preferably a DNA amplification device, in particular, Precise and quick control of multiple minute reaction solutions The present invention relates to a trace liquid reaction apparatus.
[0002]
[Prior art]
In general, various reactions using biological materials are performed under appropriate heating conditions. Recently, there has been an increasing demand for precise reaction using a small amount of biological material, and a reaction apparatus having a fine structure corresponding to the minute amount is becoming mainstream. For example, it is important that a reaction apparatus that handles a nucleic acid material such as DNA has a configuration that enables a polymerase chain reaction (hereinafter referred to as PCR) for performing DNA amplification. This PCR is a method of amplifying a target nucleic acid sequence portion from various kinds of DNA mixtures. PCR includes double stranded DNA having the DNA sequence to be amplified, two or more primers, a thermostable polymerase enzyme, and four deoxyribonucleoside triphosphates (dATP, dCTP, dGTP, dTTP). Performed in solution. The reaction consists of a heat denaturation step (about 94 ° C) to separate the double-stranded nucleic acid into a pair of single-stranded nucleic acids, and an anneal to lower the temperature and hybridize the primer to a predetermined site of the single-stranded nucleic acid. Step (approx. 50-60 ° C) and replication step of replicating a desired DNA sequence using a single-stranded nucleic acid as a template by extending the primer by conducting a polymerase reaction by raising the temperature to the thermostable enzyme operating temperature The specific nucleic acid sequence is amplified by repeating these three steps at least once or more.
[0003]
Several proposals have been made on the PCR reaction apparatus for performing the above temperature control. For example, in JP-A-5-317030, by processing a silicon wafer, forming a plurality of reaction vessels each maintained at a predetermined constant temperature, and moving the reaction solution between the plurality of reaction vessels An apparatus for performing PCR is disclosed. Also, in Japanese Translation of PCT Application Publication No. 7-508928, a silicon wafer is processed to form a reaction vessel, a resistance heating body for temperature control is formed, and the reaction solution is transported and detected automatically. Is disclosed. In these apparatuses in which a reaction vessel is formed by processing a silicon wafer, a very small amount of reaction solution is used, so that temperature control for PCR can be performed relatively accurately.
[0004]
By the way, in order to actually perform the PCR reaction, it is necessary to inject the PCR reaction solution into the reaction chamber and collect the solution after the reaction, so that the sample solution can be injected and discharged by a method such as pipetting. desirable. Also, since PCR amplifies DNA with very high sensitivity, it is desirable not to reuse the reaction vessel to prevent carryover, and for this purpose it is desirable that the reaction vessel can be easily replaced.
[0005]
Furthermore, since the reaction is carried out using a small amount of PCR solution of about several μL, loss due to evaporation of the PCR solution during the heating reaction is a big problem. In particular, if an opening for injecting and discharging the sample solution is provided, the loss due to evaporation inevitably increases.
[0006]
The above problem is a very big problem in putting a DNA amplifier into practical use, but there has been no example of researching and developing a DNA amplification device from such a practical viewpoint.
[0007]
[Problems to be solved by the invention]
In view of the above circumstances, the first object of the present invention is to use a minute reaction vessel formed on a substrate. Multiple reactions, precise and quick A reaction of a trace amount of liquid, preferably a PCR reaction, can be performed under temperature control, Very small amount The object is to provide a reaction container for a liquid, preferably a trace liquid that is easy to inject and discharge a PCR solution.
[0008]
The second object of the present invention is further: A small amount of liquid in multiple reaction chambers can be maintained at a certain level of reactivity. It is to provide a reaction vessel with a trace amount of liquid.
[0009]
The third object of the present invention is to provide a reaction container of a trace amount liquid that can easily replace the reaction container for each specimen.
[0010]
[Means for Solving the Problems]
The first problem of the present invention described above is Flat Formed on the surface of the substrate Multiple elongated channel reaction chambers And placed on the surface of the substrate Grooved reaction chamber A reaction body provided with a lid for closing the opening of the first and second through holes formed in the lid at positions corresponding to the first and second ends of each groove-shaped reaction chamber A container device; A plurality of elongated hollow spaces that are independent from each other on the same plane are provided in parallel, each having an injection port provided separately in one of the first and second through holes and a discharge port provided separately in the other. In addition, the first through hole and the second through hole have the same structure. This is achieved by a trace liquid reactor.
[0011]
The first purpose is to And an upper frame member provided so as to extend outward from the edge of the substrate; and an injection drilled in the upper frame member so as to communicate with the concave groove through the first through hole. A port; a discharge port drilled in the upper frame member so as to communicate with the concave groove through the second through hole; provided along the rear edge of the substrate and projecting outward And a module structure that is fixed by the upper frame member and the lower frame member while both surfaces of the reaction vessel device are exposed to the outside air. It is more preferably achieved by a reaction apparatus for a trace liquid characterized by the following.
[0012]
The second object of the present invention is also achieved by a trace liquid reactor having the above-described configuration.
[0013]
In addition to the above reaction vessel module, the third problem of the present invention is that And an attachment member for detachably installing the module. This is achieved by a trace liquid reactor.
[0014]
Embodiment
Hereinafter, an example in which the trace liquid reaction apparatus of the present invention is applied to a DNA amplification apparatus will be described in detail.
[0015]
FIG. 1 is a perspective view showing the overall configuration of a DNA amplification apparatus according to an embodiment of the present invention. As shown in the figure, this apparatus includes a reaction vessel module 10 having a rectangular plane, and an attachment 1 on which the module 10 is detachably installed. The attachment 1 has a base 2 and a cubic block-shaped pedestal 3 fixed thereon, and a cubic hollow storage portion 5 is formed in the center of the pedestal 3. The horizontal top surface 4 of the pedestal 3 constitutes a mounting surface for mounting the reaction vessel module 10. At the four corners of the mounting surface, bowl-shaped positioning blocks 7 are stacked. In practice, the positioning block 7 is formed integrally with the base 3. The thickness of the positioning block 7 is about ½ of the mounting surface, and the reaction container module 10 is detachably mounted in an area limited by the inner walls of the four positioning blocks. The gap between the adjacent positioning blocks 7 facilitates the handling of the module 10 by the robot arm, for example, when the reaction vessel module 10 is transported by the robot arm. A ball plunger 26 is provided on the inner wall of the positioning block 7 so that the reaction vessel module inserted from above can be fixed. Further, a pin terminal 6 is implanted on the mounting surface 4, and the pin terminal is retracted and protruded by a spring. The pin terminal 6 is disposed so as to come into contact with an electrical terminal exposed on the bottom surface of the module when the reaction container module is installed on the attachment, and to be electrically connected to an electric circuit of the reaction container module described later. ing. Although not shown, the reaction chamber module 10 is cooled by providing an electric fan in the hollow housing portion 5. Instead of the electric fan, another cooling mechanism such as a water cooling mechanism or a Peltier element may be used. Although not shown, a terminal connected to the pin terminal 6 is provided on the side surface of the base 3, and this terminal is connected to a controller (not shown) via a cable.
[0016]
As described above, in the DNA amplification device of this embodiment, since the reaction container module is detachably attached to the attachment 1, the reaction container can be easily replaced for each sample, and the effect of preventing carryover can be obtained. It is done.
[0017]
FIG. 2 is an exploded perspective view showing the structure of the reaction vessel module 10. As illustrated, the reaction vessel module 10 includes a reaction vessel device 11, an upper frame member 31, and a lower frame member 41. The reaction vessel device 11 is a plate having a rectangular plane, and eight sets of injection-side through holes 12a and discharge-side through holes 12b are formed along opposing long sides. A groove-like reaction chamber is formed between each of the eight pairs of the injection side through holes and the discharge side through holes 12a and 12b. The three elements of the injection side through hole 12a, the reaction chamber, and the discharge side through hole 12b communicate with each other, and eight sets have hollow spaces that are independent from each other.
[0018]
3A to 3C show details of the reaction vessel device. As shown in FIG. 3A, the reaction vessel device 11 includes a rectangular silicon substrate 13 and a silicon lid 15. The silicon substrate 13 and the lid 15 are cut out from a silicon wafer used for manufacturing a semiconductor device such as an IC. The dimensions of the silicon substrate 13 and the lid 15 are not particularly limited, but are 34 mm × 28 mm × 1.3 mm in this embodiment. On the surface side of the silicon substrate 13, eight grooves 14 parallel to the short side direction are formed. These grooves 14 are formed by a selective etching method well known in the semiconductor technology. On the other hand, the cover body 15 is formed with through holes 12 a and 12 b corresponding to both ends of the groove 14. Therefore, as shown in the drawing, by superimposing both, an elongated flat reaction chamber opened by the through holes 12a and 12b is formed. What is necessary is just to set the volume of the reaction chamber suitably according to the required amount of the liquid to process, and is 25 microliters in this Example.
[0019]
A temperature sensor pattern 16 and a heater pattern 17 are formed on the back side of the silicon substrate 13 as shown in FIG. A terminal 18 connected to the temperature sensor pattern 16 is formed, and a terminal 19 connected to the heater pattern 17 is formed. These terminals are formed so as to come into contact with the pin terminals 6 formed on the ground plane 4 when the reaction container module 10 is installed on the attachment 1 shown in FIG. These patterns 16 to 19 are formed by patterning a titanium thin film by a thin film forming method and photophysography well known in the semiconductor manufacturing technology. Further, as shown in FIG. 3C, these patterns 16 and 17 are formed under the groove 14 in a state of being electrically insulated from each other. That is, first, the temperature sensor pattern 16 is formed on the back surface of the silicon substrate 13, and this is covered with an interlayer insulating film 20 made of an insulating resin such as polyimide, and then the heater pattern 17 is formed. Finally, in order to improve chemical stability, an insulating resin film 21 covering the whole is formed. The heater pattern 17 is for heating the PCR sample solution in the reaction chamber (groove 14) to a temperature required for each step of the PCR, and the sensor pattern detects the temperature of the PCR sample solution in the reaction chamber. Is for.
[0020]
Next, returning to FIG. 2, the upper frame member 31 and the lower frame member 41 will be described. The upper frame member 31 has a frame shape. The material is not particularly limited, but it is desirable to use a metal material in order to cool the packing 34 to be described later and to secure a necessary strength during handling by the robot arm. In particular, iron materials such as aluminum and stainless steel, copper, brass and the like are preferable in consideration of ease of processing and high processing accuracy. In this embodiment, an aluminum upper frame member 31 is used. A recess (not shown) for fitting the reaction solution device 11 is formed on the rear surface side of the upper frame member 31. A large opening 32 is formed at the center of the upper frame member 31, and most of the reaction vessel device 11 fitted in the recess on the back surface side is exposed through the opening 32. This achieves efficient heat dissipation from the reaction vessel device. On the other hand, on the surface side of the upper frame member, an injection port 33a and a discharge port 33b penetrating the upper frame member are provided at positions corresponding to the through holes 12a and 12b of the reaction vessel device 11 fitted therein. A cylindrical packing 34 is inserted from below into each of the injection port 33a and the discharge port 33b. The size of the frame member 31 is not particularly limited, but is 45 mm × 37 mm × 9 mm in this embodiment.
[0021]
On the other hand, the lower frame member 41 is configured by combining two vertical frame members 42 and two horizontal frame members 43. The horizontal frame 43 is provided with a convex edge 44 at a position corresponding to the row of the through holes 12 a and the row of the through holes 12 b, and the convex edge 44 abuts on and supports the bottom surface of the reaction vessel device 11. Each of the vertical frames 42 is formed with two screw holes 45, and each of the horizontal frames 43 is provided with a screw hole 45 in a protruding portion 46 provided outward from the center. Then, a screw 47 is inserted into each screw hole 45 and screwed into a screw hole formed on the back surface of the upper frame member 31, whereby the reaction container device 11 is placed between the upper frame member 31 and the lower frame member. Then, the reaction vessel module is assembled. That is, the lower frame member 41 serves as a pressing plate for coupling the reaction vessel device 11 to the upper frame member 31 via the packing 34. At that time, the lower frame member 41 is in direct contact with the laminated portion of the heater pattern 17 and the temperature sensor pattern 16 on the back surface of the reaction container device. Therefore, in order not to let the heat generated in the heater pattern 17 escape, a material having a low thermal conductivity is preferable, but a material having a low thermal conductivity often has a large heat capacity. If the heat capacity is large, the heat distribution state during cooling becomes non-uniform. Therefore, in this embodiment, the lower frame member is provided with the convex edge 44 so as to minimize the contact area. Convex edge 44 of packing 34 Central axis Since it is located on the extension of, efficient fixation is possible. Although the material of the lower frame member 41 is not particularly limited, as described above, since it directly contacts the temperature sensor pattern 16 and the heater pattern 17, the thermal conductivity is low and the heating temperature range of the reaction vessel device is low. It must be stable. Examples of such a material include polycarbonate resin, epoxy resin, polyimide resin, fluorine resin, vinyl chloride resin, and ceramic material having high heat resistance. However, since the strength and altitude must be high in order to fix the reaction vessel device 11, an epoxy resin, a polyimide resin, or a ceramic material is preferable. In this example, polyimide resin was used. The size of the lower frame member 41 depends on the shapes of the upper frame member 31 and the reaction vessel device 11, and is 41.4 mm × 35.4 mm × 3 mm in this embodiment.
[0022]
Next, the structure of the injection port 33a and the discharge port 33b in the reaction container module assembled as described above will be described with reference to FIG. The silicon substrate 13 and the lid 15 sandwiched between the upper frame member 31 and the lower frame member 43 define a reaction chamber composed of the groove 14. As shown, the lower surface of the silicon substrate contacts the lower frame member 43 only at the convex edge 44 of the lower frame member. As shown in the drawing, the through-hole of the injection port 33a or the discharge port 33b has an inlet portion having a bowl-shaped cross-sectional shape, and has an open end portion that is widest and tapers toward the narrowest neck portion. The diameter of the neck portion is slightly larger than the pipette tip tip outer diameter described later. When the pipette tip tip outer diameter is 0.8 mm, about 1.0 mm is preferable. The central portion located immediately below the neck portion has substantially the same diameter as the open end portion, and the lowermost leg portion has a wider diameter than the central portion. Accordingly, the through-hole forming the injection port or the discharge port has a first step portion between the neck portion and the central portion and a second step portion between the center portion and the leg portion. And the cylindrical packing 34 inserted from the lower side of the through hole is fixed in contact with the first step portion of the injection port through hole at its upper end. As shown in FIGS. 5A and 5B, the upper end of the cylindrical packing 34 is closed and the lower end is opened. A cross-shaped cut 35 is formed in the closed upper end surface. A convex edge 36 protruding outward is formed at the lower end of the packing 35, and this convex edge 35 is fixed in contact with the second step portion of the injection port through hole. The inner diameter of the packing 34 is substantially the same as the neck portion of the injection port through-hole and the through-hole 12a of the lid body 15, but the lower end thereof has an inner surface that is tapered outward as shown in the figure. Is spreading. However, it does not expand until the inner diameter becomes equal to the outer diameter, and the packing 34 has a flat lower end surface. Therefore, since the diameter of the lower end opening of the packing 34 is larger than the diameter of the through hole 12a formed in the lid body 15, an alignment margin between the packing 34 and the through holes 12a and 12b can be taken. However, in order to increase the contact area between the packing 34 and the lid 15 of the reaction vessel device, it may be a straight tube instead of a tapered shape. Further, since the lower end of the packing 34 protrudes downward from the lower end of the injection port through hole, only the flat lower end surface of the packing 34 contacts the top surface of the lid 15 of the reaction vessel device, and the upper frame 31 is a lid. Do not touch the body 15. The material of the packing 34 is not particularly limited, but a rubber-based resin having a high packing effect is preferable. In particular, butyl rubber and silicone rubber that are highly molded are suitable. In this example, silicone rubber was used. In this embodiment, the packing 34 has a length of 2.6 mm, a maximum outer diameter of 2.6 mm, a minimum outer diameter of 2 mm, and an inner diameter of 1 mm. A notch of mm is formed in a cross shape.
[0023]
FIG. 6 shows an example of a circuit configuration in the DNA amplification device of the above embodiment. A controller 60 is connected to the attachment 1. The controller includes a power supply 61 connected to the heater pattern 17 of the reaction vessel device, a power supply 62 connected to a cooling fan incorporated in the attachment 1, and a digital multiplier 63 connected to the temperature sensor pattern 16 of the reaction vessel device. It is equipped with. These are all connected to the personal computer 64 and controlled so as to realize a predetermined temperature cycle of the PCR reaction performed in the reaction vessel device.
[0024]
Next, the operation of the DNA amplification apparatus of the above embodiment will be described.
[0025]
In order to perform DNA amplification using this apparatus, first, the same or different kinds of PCR solutions containing necessary components are injected into each reaction chamber 14 of the reaction vessel device. The injection amount may be an amount that completely fills the inside of the reaction chamber 14, but is preferably an amount in which some air remains in the vicinity of the through holes 12a and 12b, and is 10 to 25 μL in this embodiment. At the time of injection, as shown in FIG. 6, the pipette tip 50 is inserted into an injection port 33a formed in the upper frame member 31 of the reaction container module. The pipette tip 50 enters by pushing open a cross-shaped cut 35 formed on the closed top surface of the packing 34 and is fixed in the state shown in the figure. At this time, the side surface of the pipette tip 50 is in good contact with the cut portion 35 of the packing 34, so that the PCT solution can be completely injected and discharged. Further, as compared with the case where the packing 34 is not present, sufficient adhesion can be obtained without pressing the pipette tip, so that the pipette tip can be easily inserted and removed. When the PCR sample solution is injected and discharged from one of the injection and discharge ports 33a and 33b, the cut 35 of the packing 34 of the other port is slightly opened upward elastically by the pressurized air. Can go in and out. This prevents extraction of the pipette tip after injection, insertion of the pipette tip at the time of discharge, or scattering of the liquid during injection. As a result, the risk of infection is eliminated, and the pressure in the reaction vessel during injection and discharge is adjusted by smooth insertion and removal of the pipette tip and gentle and smooth movement of the liquid. Done. At this time, when the air is insufficiently entered and exited, a small through hole may be formed at the center of the cross-shaped cut portion 35. Moreover, you may open the top part of packing completely. Since the lower ends of the walls of the injection and discharge ports 33a and 33b are sealed by the protruding portion of the packing 34 between the bottom of the reaction vessel device and the lid 15 of the reaction vessel device, the liquid between them is sealed. There will be no leakage. Since the bottom surface of the protruding portion of the packing 43 is flat, the sealing property between the top surface of the lid 15 is good.
[0026]
When the PCR sample solution is injected into the reaction chamber in this way, the reaction container module 10 is set in the attachment 1 as shown in FIG. 1, and the controller 60 in FIG. 6 is operated. However, the PCR sample solution may be injected after the reaction container module 10 is installed in the attachment 1. The power supply 61 of the controller 60 supplies current to the heater pattern 17 formed on the back surface of the reaction vessel device 11 according to a preset temperature cycle program to generate heat, and heats the PCR sample solution in the reaction vessel. At this time, vapor is generated from the sample solution. As can be seen from FIG. 4, the top surface of the packing 34 is closed, so that the dissipation of the vapor to the outside is limited. Further, since the packing 34 is sufficiently cooled by the upper frame member 31, most of the vapor condenses inside the packing 34, and therefore does not leak to the outside. In addition, in the vicinity of the injection-side through-hole 12a and the vicinity of the discharge-side through-hole 12b, there is a slight amount of air that is not replaced by the PCR solution filled in the groove 14 as a reaction chamber. This air is also cooled by the packing 34. As a result, excessive volume expansion is unlikely to occur, and liquid splash and vapor escape from the reaction chamber are effectively prevented. Furthermore, since both the injection-side through-hole 12a and the discharge-side through-hole 12b have the same structure, the pressure on the injection side and the discharge side is uniform even at high temperatures. Since it is held near the center of the reaction chamber 14, there is an advantage that the reactivity can be kept constant. In addition, as can be seen from FIG. 4, the reaction vessel device 11 is in contact with the upper frame member 31 only at its side end surface, and its top surface (that is, the lid 15) is in contact only with the packing 34 to be in contact with the upper frame member 31. There is no contact. Accordingly, it is possible to prevent the temperature distribution from becoming non-uniform due to the heat of the reaction vessel escaping through the upper frame member 31.
[0027]
In the temperature reduction process of the PCR cycle, the power supply 61 is turned off under the control of the personal computer 64, the power supply 63 is turned on, and air cooling by a fan is performed. At this time, since a considerable area of the front surface and the back surface of the reaction container device 11 is exposed, efficient cooling is achieved.
[0028]
Thus, using the DNA amplification apparatus of the above example, the temperature cycle test in which the reaction vessel device 11 was filled with the PCR sample solution and maintained at the temperatures of 68 ° C. and 92 ° C. was repeated, and the results shown in FIG. It was. As can be seen from this result, it was possible to achieve very accurate temperature control.
[0029]
As described above, the top surface of the packing 34 may be opened in some cases, but in that case, loss due to evaporation of the PCR sample solution in the reaction chamber becomes a problem. Therefore, as a countermeasure in this case, as shown in FIG. 9, an evaporation prevention lid 70 for closing the injection port and the discharge ports 33 a and 33 b is placed on the upper frame member 31. preferable. As this evaporation prevention lid, for example, a metal plate can be used. Further, in order to position the evaporation preventing lid 70, positioning blocks 38 are protruded from the four corners of the upper frame member 31, and cutouts 72 corresponding to the positioning blocks 38 are formed at the four corners. Thereby, the protrusion 71 of the lid 70 is placed and fixed on the surface of the upper frame member between the blocks 38, and closes the injection port and the discharge ports 33a and 33b. Further, it is preferable that the protruding portion of the lid 70 protrudes to the outside of the upper frame member 31 in order to facilitate handling by the robot arm. Further, for example, by making the top surface level on the short side facing the upper frame member lower than the top surface level on the long side, a gap into which the robot arm can be inserted is formed between the lid 70 and the placement surface. You may make it do. Further, an accurate and continuous high-speed system may be designed by performing three-dimensional movement of the pipette tip and liquid suction / discharge operation, or monitoring the temperature distribution in the reaction chamber with a sensor. Further, the reaction container module may be repeatedly reused by performing appropriate and sufficient cleaning or sterilization treatment, so that the module may not be replaced.
[0030]
【The invention's effect】
As detailed above, according to the present invention, Multiple slender reaction chambers are independently formed in a small reaction vessel device, so precise and quick A PCR reaction can be performed under temperature control, each PCR to reaction vessel for It is possible to provide a DNA amplification device that can easily inject and discharge a solution.
[0031]
Moreover, according to the present invention, In the slender groove-like reaction chamber, the air pressure on the injection port side and the discharge port side becomes uniform even at high temperatures, so even if the amount is small, the PCR solution is held near the center of the reaction chamber and the reaction is kept constant. Can be kept in .
[Brief description of the drawings]
FIG. 1 is an exploded perspective view showing the overall configuration of a DNA amplification apparatus according to an embodiment of the present invention.
2 is an exploded perspective view showing details of a reaction vessel module 10 in FIG. 1. FIG.
3 is an exploded perspective view showing a semiconductor substrate and a lid constituting the reaction vessel (A) in FIG. 2. FIG. (B) is a top view which shows the back surface of a semiconductor substrate. (C) is sectional drawing which shows the groove part of a semiconductor substrate.
4 is an enlarged cross-sectional view showing an injection port portion of the reaction vessel module of FIG. 2;
5A is a plan view of the packing, and FIG. 5B is a front view thereof.
FIG. 6 is a diagram showing an electric circuit configuration of the DNA amplification device according to the embodiment of FIGS.
FIG. 7 is a cross-sectional view showing a state in which a PCR sample solution is injected into a reaction container device.
FIG. 8 is a graph showing the results of a temperature cycle test of the DNA amplification apparatus according to the example of FIGS.
FIG. 9 is an exploded perspective view showing another embodiment of the DNA amplification device according to the present invention.
[Explanation of symbols]
DESCRIPTION OF SYMBOLS 1 ... Attachment, 2 ... Base, 3 ... Base, 4 ... Top surface of base, 5 ... Hollow storage part, 6 ... Pin terminal, 7 ... Positioning block 7, 10 ... Reaction container module, 11 ... Reaction container device, 12a ... injection-side through-holes 12a, 12b ... discharge-side through-holes, 13 ... semiconductor substrate, 14 ... groove, 15 ... lid, 16 ... temperature sensor pattern, 17 ... heater pattern, 18, 19 ... terminal, 20, 21 ... polyimide Membrane, 31 ... upper frame member, 32 ... opening, 33a ... injection port, 33b ... discharge port 33b, 34 ... packing 34, 35 ... cross notch, 36 ... convex edge, 38 ... positioning block, 41 ... lower frame Member, 42 ... Vertical frame member, 43 ... Horizontal frame member 43, 44 ... Convex edge 44, 45 ... Screw hole, 47 ... Screw, 50 ... Pipette tip, 60 ... Controller, 61 ... Power source, 62 ... Power source, 63 ... Multi Laiya, 70 ... anti-evaporation cover, 71 ... protruding portion, 72 ... notch

Claims (7)

A plurality of elongated groove-like reaction chambers formed on the surface of the flat substrate, a lid placed on the surface of the substrate and closing an opening of the groove-like reaction chamber , and each groove-like reaction chamber A reaction vessel device having first and second through holes formed in the lid at positions corresponding to the first and second ends of the reaction chamber; one of the first and second through holes ; A plurality of elongated hollow spaces arranged in parallel to each other on the same plane, and the first through holes and the first through holes. A trace liquid reactor with the same structure of the two through holes . 2. The trace liquid reaction apparatus according to claim 1, wherein cooling members for accelerating cooling are arranged for all the injection ports and the discharge ports . 2. The trace liquid reaction apparatus according to claim 1 , further comprising an upper frame member provided so as to extend along an edge portion of the substrate and to the outside thereof; and the concave groove through the first through hole. An injection port drilled in the upper frame member to communicate with the upper frame member; a discharge port drilled in the upper frame member to communicate with the concave groove through the second through hole; and the substrate And a lower frame member provided so as to extend outward from the rear edge of the container, and is fixed by the upper frame member and the lower frame member while both surfaces of the reaction container device are exposed to the outside air. A trace liquid reaction apparatus characterized by that . 4. The trace liquid reaction apparatus according to claim 3, wherein the lower frame member has a structure in contact with the reaction vessel device through a convex low thermal conductivity member. apparatus. 4. The trace liquid reaction apparatus according to claim 3 , further comprising an attachment member for detachable installation . The apparatus according to any one of claims 1 to 5, further comprising a temperature sensor pattern and a heater pattern . 6. The apparatus according to claim 5 , further comprising connection means for electrically connecting a temperature sensor pattern and a heater pattern provided in the reaction device to an external controller. apparatus.

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