Patent Publication Number: US-2005123457-A1

Title: Reaction vessel and reaction apparatus

Description:
TECHNICAL FIELD  
      This invention relates to a reaction vessel, reaction apparatus, and method with which the temperature of a reaction solution can be rapidly controlled and the reaction can be monitored in real time.  
     BACKGROUND ART  
      A polymerase chain reaction (hereafter referred to as “PCR”) is a technique which amplifies target nucleic acids by raising and lowering the temperature of a heat-resistant polymerase and primers. This technique is widely used in fields such as genetic engineering and biological test methods and detection methods.  
      The principle behind PCR lies in the fact that target DNA is amplified in a geometrical progression by numerous iterations of a cycle according to a thermal profile (rise and fall of temperature) that is set in three stages: a first stage in which the temperature is maintained at a level at which double-stranded DNA containing a target DNA sequence dissociates into a single strand, a second stage in which the temperature is maintained at a level at which forward and reverse primers are annealed with the dissociated single-stranded DNA, and a third stage in which the temperature is maintained at a level at which a complementary DNA chain is synthesized with the single-stranded DNA by the DNA polymerase.  
      For example, a PCR can be conducted by reacting a reaction solution containing double-stranded DNA that includes a target DNA sequence, an excess amount of a pair of primers, and a heat-resistant polymerase for 30 to 40 cycles, with each cycle comprising 30 seconds at 95° C., 30 seconds at 65° C., and 1 minute at 72° C. At 95° C., the double-stranded DNA dissociates into single-stranded DNA. Next, the reaction solution is cooled to an appropriate temperature as dictated by the base sequences of the primers (65° C. in the above example), whereupon the primers and the single-stranded DNA are annealed. The temperature is then raised to the reaction temperature of the polymerase (72° C. in the above example), whereupon a DNA synthesis reaction proceeds under the influence of the polymerase.  
      Thus, controlling the temperature of the reaction solution is important in a PCR, so a PCR is usually conducted using a thermostat apparatus that allows programming of the temperature control, and a reaction vessel that can be used with such an apparatus.  
      The most common approach is to use an apparatus in which micro-tubes are snugly fitted in holes of a metal block equipped with a heating/cooling apparatus, and a cycle of heating (dissociation of the double-stranded DNA), cooling (annealing of the primers), and heating (chain extension reaction by the polymerase) is repeated for the reaction solution in the micro-tubes via the metal block. Two different systems are employed for cooling the metal block: using a compressor, and using a Peltier cooling system. Recently, apparatuses have also been available in which the micro-tubes are moved together in their rack, rather than using a metal block, and in which the micro-tubes are successively immersed in three liquid-phase or solid-phase incubators with independent temperatures, so that a cycle consisting of heating (dissociation of the double-stranded DNA), cooling (annealing of the primers), and heating (chain extension reaction by the polymerase) is repeated.  
      If a large number of specimens is involved, in order to process numerous specimens all at once, as when a PCR is conducted for the purpose of screening, apparatuses have been developed with which PCRs for 96 specimens can be conducted at the same time using a PCR micro-titer plate (96 wells).  
      In particular, there has been a growing need for the efficient processing of numerous specimens in parallel by automating a series of operations comprising the preparation of samples containing target nucleic acids (such as extraction of nucleic acids from cells), amplification of these target nucleic acids by PCR, and monitoring of the progress of the PCR (such as whether or not the target nucleic acids have been amplified, or the amount of PCR amplification product), in order to treat numerous specimens more efficiently in genetic diagnosis and the genome project. If this series of operation is to be automated and numerous specimens are to be efficiently processed in parallel, it is necessary first of all to minimize the time PCR takes, secondly to minimize the quantity of specimen required for a PCR, and thirdly to monitor the progress of the PCR in real time (that is, instantly during the course of the PCR).  
      However, with a conventional PCR reaction apparatus and PCR reaction vessel, since the object is to perform a PCR by means of typical temperature control in which a reaction is repeated for 30 to 40 cycles, with each cycle comprising 30 seconds at 95° C., 30 seconds at 65° C., and 1 minute at 72° C., it is difficult to achieve the goal of minimizing the time required for the PCR by using a conventional PCR reaction apparatus and PCR reaction vessel. For example, when a reaction is repeated for 30 to 40 cycles using a conventional PCR reaction apparatus and PCR reaction vessel with each cycle comprising 30 seconds at 95° C., 30 seconds at 65° C., and 1 minute at 72° C., it takes about 1 hour to complete the PCR.  
      Also, with a conventional PCR reaction apparatus and PCR reaction vessel, if the amount of specimen (reaction solution) is too small, the solvent (ordinarily water) in the reaction solution may evaporate during the PCR, bringing the reaction to a halt. The reasons for this include the following. Because of the large contact area between air and the reaction solution in the reaction chamber (such as a micro-tube or micro-titer plate well) in which the PCR proceeds, the solvent in the reaction solution is in an environment in which it is prone to evaporation, and since the temperature of the walls inside the reaction chamber is not uniform, some portions of the walls inside the reaction chamber are lower in temperature than the reaction solution (such as the upper part of a micro-tube or upper part of a micro-titer plate well), so the evaporated solvent ends up being liquefied in these areas. Accordingly, it is difficult to achieve the goal of minimizing the amount of reaction solution by using a conventional PCR reaction apparatus and PCR reaction vessel.  
      In light of this situation, an apparatus has been developed in which a small amount of reaction solution is enclosed inside a micro-capillary which has a large surface area and good thermal conductivity, and heating and cooling are performed by means of hot air from a halogen lamp or other such heat source and room-temperature cool air. For example, LightCycler (made by Roche Molecular Biochemicals) is marketed as an apparatus of this type. With this apparatus, temperature control of approximately 20° C./sec can be achieved by utilizing micro-capillaries that have a large surface area and a good thermal conductivity. Thus, each cycle takes only about 30 to 60 seconds, so 30 cycles can be completed in about 15 to 30 minutes. Also, since micro-capillaries are utilized, a PCR can be conducted using a very small amount of reaction solution, only about 5 to 20 μl. Furthermore, since one of the characteristics of a glass capillary is that it focuses nearly all the irradiation light at the tip of the capillary, fluorescent light emitted from a reaction solution according to the amount of PCR amplification product can be measured quickly and with good sensitivity, making it possible to monitor the progress of the PCR in real time.  
      Thus, a PCR reaction apparatus that makes use of a micro-capillary as a PCR reaction vessel reduces the time the PCR takes by means of rapid temperature control of the reaction solution, and reduces the amount of reaction solution required for the PCR to an extremely small amount. Furthermore, the progress of the PCR can be monitored in real time. Such a PCR reaction apparatus is therefore extremely useful when a PCR is conducted alone.  
      With this PCR reaction apparatus, however, the filling of the micro-capillaries with the reaction solution requires an operation in which the reaction solution is added to plastic containers disposed at the upper parts of glass capillaries, and sealed in with plastic stoppers, after which a centrifuge is used to move the reaction solution from the plastic containers into the glass capillaries, and the various capillaries are then removed from the centrifuge and placed in the reaction apparatus. Also, if air is admixed in the course of filling the micro-capillaries, this air will expand as a result of the heating performed in the process of the PCR, causing the reaction solution to move through the micro-capillaries and resulting in a drop in the amplification efficiency of the PCR. Consequently, great care must be exercised in the filling of the micro-capillaries with the reaction solution.  
      Therefore, it is difficult to utilize a PCR reaction apparatus in which micro-capillaries are used as the PCR reaction vessel for the automation of the series of operations comprising the preparation of samples containing target nucleic acids (such as extraction of nucleic acids from cells), the amplification of the target nucleic acids by PCR, and monitoring the progress of the PCR (such as whether or not the target nucleic acids have been amplified, or the amount of PCR amplification product).  
      Also, whichever PCR reaction apparatus and PCR reaction vessel are used, the reaction vessel main body holding the reaction solution must be covered with a lid and the inside of the reaction chamber (such as microtubes or the wells of a microtiter plate) in which the PCR proceeds must be sealed in order to prevent the reaction from coming to a halt when the solvent (usually water) in the reaction solution evaporates in the middle of the PCR. Therefore, with a conventional PCR reaction apparatus and PCR reaction vessel, accessing the amplified fragments obtained by PCR first requires that the lid be removed from the reaction vessel main body, so it was difficult to automate the work from the amplification of the target nucleic acids by PCR up to the accessing of the amplified fragments.  
     DISCLOSURE OF THE INVENTION  
      In view of this, a first object of the present invention is to provide a reaction vessel, reaction apparatus, and method with which a reaction can be automated without requiring centrifugation when a reaction solution is held in a reaction chamber, the temperature of the reaction solution held in the reaction chamber can be rapidly controlled, the reaction can proceed even when just a tiny amount of reaction solution is held in the reaction chamber, and the reaction occurring in the reaction chamber can be monitored in real time (that is, instantly during the course of the reaction).  
      A second object of the present invention is to provide a reaction vessel, reaction apparatus, and method with which the above-mentioned first object can be achieved, and after the reaction has been conducted with the reaction vessel main body covered by the cover member, the reaction product contained in the reaction solution inside the reaction vessel can be accessed without removing the cover member from the reaction vessel main body.  
      (1) In order to achieve the stated objects, the reaction vessel of the present invention comprises a reaction vessel main body equipped with a reaction chamber having an opening and capable of holding a reaction solution, and a cover member capable of sealing the opening of the reaction chamber, wherein the cover member and the reaction chamber have a contact surface that comes into contact with the reaction solution held in the reaction chamber in a state in which the cover member is mounted on the reaction vessel main body, and the cover member is made of a light transmitting material so that light can pass from the reaction solution held in the reaction chamber, through the contact surface of the cover member, to the outside of the reaction vessel, or the reaction vessel main body is made of a light transmitting material so that light can pass from the reaction solution held in the reaction chamber, through the contact surface of the reaction chamber, to the outside of the reaction vessel.  
      The reaction chamber provided to the reaction vessel main body has an opening and is capable of holding a reaction solution, and the reaction solution is added through the opening in the reaction chamber and held in the reaction chamber. The reaction chamber is the place where the desired reaction takes place, and components required for the desired reaction to take place, reagents required for the measurement of the reaction progress (such as fluorescent dyes and other light-emitting substances), and so forth are contained in the reaction solution held in the reaction chamber.  
      There are no particular restrictions on the structure of the reaction chamber, as long as it has an opening and is capable of holding a reaction solution. In general, the reaction chamber is formed in the reaction vessel main body as a concave component having an opening at its upper end. The reaction chamber is preferably formed on the reaction vessel main body as a concave component consisting of a thin plate. In this case, heat movement between the outside of the reaction chamber and the reaction solution inside the reaction chamber occurs through this thin plate, allowing the temperature of the reaction solution to be controlled more rapidly and efficiently. Also, irradiation conditions and light reception conditions can be set more easily if the irradiation of the reaction chamber with light and the detection of light emitted from the reaction solution are performed through this thin plate.  
      There is no need for the reaction chamber to have a capillary structure, nor is it essential for centrifuging to be performed when the reaction solution is held in the reaction chamber. The reaction vessel of the present invention is designed such that when the cover member is placed over the reaction vessel main body, the cover member (e.g. a convex component provided to the cover member) enters the interior of the reaction chamber through the opening in the reaction chamber, and comes into contact with the reaction solution held in the reaction chamber. It is therefore preferable for the reaction chamber to have a structure that allows easy entry of cover member (such as a convex component provided to the cover member). It is also preferable, from the standpoint of automating the aliquoting of the reaction solution, for the reaction chamber to have a structure that allows the reaction solution added through the opening to reach the bottom of the reaction chamber without any downward force other than gravity being applied to the reaction solution. Accordingly, it is actually inappropriate for the reaction vessel of the present invention to have a reaction chamber with a capillary-like structure.  
      When the cover member covers the reaction vessel main body, the opening of the reaction chamber is sealed off by the cover member. This prevents the reaction solution held in the reaction chamber from being contaminated, and allows the desired reaction to be conducted more accurately within the reaction chamber. If the reaction vessel main body is equipped with a plurality of reaction chambers, then the openings of the various reaction chambers can be sealed by the cover member so as to prevent admixture of the reaction solution contained in one reaction chamber with the reaction solution contained in other reaction chambers, which allows the desired reaction to be accurately conducted in the various reaction chambers.  
      In terms of preventing contamination of the reaction solution contained in the reaction chamber, it is preferable for the cover member to have a first snug-fit component capable of fitting snugly against the periphery of the opening in the reaction chamber. In this case, the first snug-fit component of the cover member and the periphery of the opening in the reaction chamber fit snugly together to seal the reaction chamber, which prevents contamination of the reaction solution.  
      Also, in terms of preventing contamination of the reaction solution contained in the reaction chamber, it is preferable for the cover member to have a second snug-fit component capable of fitting snugly against the inner peripheral surface of the reaction chamber (the inner peripheral surface of the concave component formed in the reaction vessel main body). In this case, the second snug-fit component of the cover member and the inner peripheral surface of the reaction chamber fit snugly together to seal the reaction chamber, which prevents contamination of the reaction solution.  
      There are no particular restrictions on the structure of the cover member, as long as it has a contact surface capable of coming into contact with the reaction solution when the cover member is mounted on the reaction vessel main body. An example of the cover member structure is a flat plate in which a convex component has been formed. In this case, when the cover member is placed over the reaction vessel main body, the convex component enters the interior of the reaction chamber through the opening in the reaction chamber, and comes into contact with the reaction solution held in the reaction chamber. The convex component formed in the cover member preferably comprises a thin plate. In this case, heat movement between the outside of the reaction chamber and the reaction solution inside the reaction chamber occurs through this thin plate, allowing the temperature of the reaction solution to be controlled more rapidly and efficiently. Also, irradiation conditions and light reception conditions can be set more easily if the irradiation of the reaction chamber with light and the detection of light emitted from the reaction solution are performed through this thin plate.  
      The cover member and the reaction chamber have a contact surface that comes into contact with the reaction solution held in the reaction chamber when the cover member is mounted on the reaction vessel main body. In the present invention, the surface of the reaction chamber that comes into contact with the reaction solution is called the “contact surface of the reaction chamber,” and the surface of the cover member that comes into contact with the reaction solution is called the “contact surface of the cover member.” The contact surface of the reaction chamber and the contact surface of the cover member do not necessarily refer to a specific surface, and will vary (for example, increase and decrease the contact area) with the conditions (for example, the volume of reaction solution held in the reaction chamber). For instance, when the convex component of the cover member presses against the reaction solution, it raises the level of the reaction solution, which increases the contact surface of the reaction chamber and the contact surface of the cover member.  
      With the reaction vessel of the present invention, the temperature of the reaction solution is controlled as necessary during the desired reaction inside the reaction chamber. The temperature of the reaction solution is usually controlled after the cover member has been placed over the reaction vessel main body. However, reaction solution temperature control may also be carried out before the cover member is placed over the reaction vessel main body and/or during the process of the placing the cover member over the reaction vessel main body. If control of the reaction solution temperature is performed after the cover member has been placed over the reaction vessel main body, the temperature of the reaction solution can be controlled by the movement of heat through the contact surface of the reaction chamber and/or the contact surface and the cover member. This allows the temperature of the reaction solution to be controlled rapidly.  
      With the reaction vessel of the present invention, there are no particular restrictions on the reaction occurring within the reaction chamber, but the reaction vessel of the present invention can be used to advantage in reactions that demand control of the reaction solution temperature when the reaction is commenced, during its progress, or when it is halted (such as an enzyme reaction), and is especially suitable for use in reactions in which the temperature of the reaction solution needs to be controlled periodically or over time during the course of the reaction (such as a PCR). The phrase “control of the reaction solution temperature” as used here refers both to varying (raising and lowering) the temperature of the reaction solution and to maintaining the temperature of the reaction solution.  
      The reaction vessel of the present invention may further comprise a heat-conducting metal block or heat-conducting metal plate provided so as to be in contact with the reaction vessel main body and/or the cover member. In this case, temperature control of the reaction vessel main body is performed through the contact surface between the reaction vessel main body and the heat-conducting metal block or heat-conducting metal plate, and temperature control of the cover member is performed through the contact surface between the cover member and the heat-conducting metal block or heat-conducting metal plate. Temperature control of the reaction solution, meanwhile, is performed through the contact surface of the reaction chamber and/or the contact surface of the cover member. The heat-conducting metal block or heat-conducting metal plate may be provided so as to be in contact with either the reaction vessel main body or the cover member, or so as to be in contact with both. Since the heat-conducting metal block or heat-conducting metal plate can easily be molded to conform to the shapes of the reaction vessel main body and cover member, the contact area with the reaction vessel main body and the cover member can be increased. As a result, heat can be moved efficiently via the heat-conducting metal block or heat-conducting metal plate. In addition to being used as a medium for the movement of heat (heat exchanger), the heat-conducting metal block or heat-conducting metal plate can also be used as a member that supports the reaction vessel main body, or as a member that applies pressure to the cover member when the cover member is placed over the reaction vessel main body.  
      With the reaction vessel of the present invention, when the cover member is placed over the reaction vessel main body, the cover member (e.g. a convex component provided to the cover member) enters the interior of the reaction chamber through the opening in the reaction chamber, so that the air or other gas present inside the reaction chamber is pushed out of the reaction chamber, and the opening of the reaction chamber is sealed in this state. Therefore, the amount of air or other gas present inside the reaction chamber is less than that prior to the covering of the reaction chamber by the cover member. Furthermore, since the cover member (e.g. a convex component provided to the cover member) that enters the interior of the reaction chamber comes into contact with the reaction solution held in the reaction chamber, the contact area between the reaction solution and the air or other gas present in the reaction chamber is less than that prior to the covering by the cover member. Thus, when the cover member is placed over the reaction vessel main body, there is less air or other gas present inside the reaction chamber, and the contact area between the reaction solution and the air or other gas present inside the reaction chamber is also reduced, so when the desired reaction is conducted inside the reaction chamber, the evaporation of the reaction solution into the air or other gas present inside the reaction chamber can be suppressed. As a result, the reaction can proceed even when just a tiny amount of reaction solution is held in the reaction chamber.  
      With the reaction vessel of the present invention, the cover member is made of a light transmitting material so that light can pass from the reaction solution held in the reaction chamber, through the contact surface of the cover member, to the outside of the reaction vessel, or the reaction vessel main body is made of a light transmitting material so that light can pass from the reaction solution held in the reaction chamber, through the contact surface of the reaction chamber, to the outside of the reaction vessel.  
      The structure may be such that the light going from the reaction solution held in the reaction chamber to the outside of the reaction vessel can be transmitted through part of the contact surface of the cover member, or can be transmitted through all of the contact surface of the cover member. Also, as long as light can be transmitted from the reaction solution held in the reaction chamber, through the contact surface of the cover member, to the outside of the reaction vessel, the structure may be such that only part of the cover member is made of a light transmitting material, or such that all of the cover member is made of a light transmitting material.  
      The structure may be such that the light going from the reaction solution held in the reaction chamber to the outside of the reaction vessel can be transmitted through part of the contact surface of the reaction chamber, or can be transmitted through all of the contact surface of the reaction chamber. Also, as long as light can be transmitted from the reaction solution held in the reaction chamber, through the contact surface of the reaction chamber, to the outside of the reaction vessel, the structure may be such that only part of the reaction vessel main body is made of a light transmitting material, or such that all of the reaction vessel is made of a light transmitting material.  
      With the reaction vessel of the present invention, just the cover member or the reaction vessel main body may be made from a light transmitting material, or both may be made from a light transmitting material. If just the cover member or the reaction vessel main body is made from a light transmitting material, then the other will be made of an opaque material.  
      There are no particular restrictions on the type of light transmitting material, and any material can be used that is transparent or semi-transparent and has the strength required of the cover member and the reaction vessel main body. Examples of this material include plastics and glass.  
      With the reaction vessel of the present invention, the light (such as fluorescent light or chemical luminescence) emitted from the reaction solution held in the reaction chamber is transmitted through the contact surface of the cover member and/or the contact surface of the reaction chamber to the outside of the reaction vessel. Specifically, with the reaction vessel of the present invention, the light (such as fluorescent light or chemical luminescence) emitted from the reaction solution held in the reaction chamber can be detected outside the reaction vessel, with the reaction solution still held inside the reaction chamber. Therefore, if the light emitted from the reaction solution is used as an index of the progress of the reaction occurring in the reaction chamber, the progress of the reaction can be monitored in real time (that is, instantly during the course of the reaction) by detecting the light emitted from the reaction solution.  
      The term “monitor” as used here includes quantitative and qualitative measurement and analysis performed continuously or intermittently during the course of the reaction, as well as quantitative and qualitative measurement and analysis after the reaction has reached a steady state or after completion of the reaction, for example. Also, the phrase “progress of the reaction” as used here includes status and degree of the reaction.  
      The light emitted from the reaction solution may be detected through just the contact surface of the cover member or the contact surface of the reaction chamber, or through both of these. The light emitted from the reaction solution may also be detected through all or part of the contact surface of the cover member and/or the reaction chamber.  
      (2) In a first aspect of the present invention, the cover member is made of a light transmitting material so that light can pass from the outside of the reaction vessel, through the contact surface of the cover member, to the reaction solution held in the reaction chamber, or the reaction vessel main body is made of a light transmitting material so that light can pass from the outside of the reaction vessel, through the contact surface of the reaction chamber, to the reaction solution held in the reaction chamber.  
      With the reaction vessel pertaining to this aspect, if the cover member is made of a light transmitting material so that light can pass from the outside of the reaction vessel, through the contact surface of the cover member, to the reaction solution held in the reaction chamber, the reaction solution held in the reaction chamber can be irradiated through the contact surface of the cover member with excitation light emitted from a laser or other light source provided to the outside of the reaction vessel of the present invention. If a fluorescent dye or other fluorescent material is added ahead of time to the reaction solution, then the fluorescent material will be excited by the irradiation of the reaction solution with the excitation light, and fluorescent light will be emitted from the reaction solution. Since the fluorescent light emitted from the reaction solution is transmitted through the contact surface of the cover member and/or the contact surface of the reaction chamber to the outside of the reaction vessel of the present invention, this light can be detected by a fluorescent light detector provided to the outside of the reaction vessel of the present invention.  
      With the reaction vessel pertaining to this aspect, if the reaction vessel main body is made of a light transmitting material so that light can pass from the outside of the reaction vessel, through the contact surface of the reaction chamber, to the reaction solution held in the reaction chamber, the reaction solution held in the reaction chamber can be irradiated through the contact surface of the reaction chamber with excitation light emitted from a laser or other light source provided to the outside of the reaction vessel of the present invention. If a fluorescent dye or other fluorescent material is added ahead of time to the reaction solution, then the fluorescent material will be excited by the irradiation of the reaction solution with the excitation light, and fluorescent light will be emitted from the reaction solution. Since the fluorescent light emitted from the reaction solution is transmitted through the contact surface of the cover member and/or the contact surface of the reaction chamber to the outside of the reaction vessel of the present invention, this light can be detected by a fluorescent light detector provided to the outside of the reaction vessel of the present invention.  
      With the reaction vessel pertaining to this aspect, the structure may be such that light going from the outside of the reaction vessel to the reaction solution held in the reaction chamber can be transmitted through part of the contact surface of the cover member and/or the reaction chamber, or can be transmitted through all of the contact surface. Also, as long as light can be transmitted from the outside of the reaction vessel, through the contact surface of the cover member, to the reaction solution held in the reaction chamber, the structure may be such that only part of the cover member is made of a light transmitting material, or such that all of the cover member is made of a light transmitting material. Also, as long as light can be transmitted from the outside of the reaction vessel, through the contact surface of the reaction chamber, to the reaction solution held in the reaction chamber, the structure may be such that only part of the reaction vessel main body is made of a light transmitting material, or such that all of the reaction vessel main body is made of a light transmitting material.  
      With the reaction vessel pertaining to this aspect, the irradiation of the reaction solution with the excitation light and the detection of the fluorescent light emitted from the reaction solution held in the reaction chamber can be performed outside the reaction vessel, with the reaction solution still held inside the reaction chamber. Therefore, if a fluorescent material that can serve as an index of the progress of the reaction is added ahead of time to the reaction solution, the progress of the reaction can be monitored in real time (that is, instantly during the course of the reaction) by detecting the fluorescent light emitted by this fluorescent material during the course of the reaction.  
      The fluorescent material that can serve as an index of the reaction progress can be suitably selected according to the type of reaction occurring in the reaction chamber. For instance, if the reaction occurring in the reaction chamber is a PCR, it is possible to use a fluorescent material whose fluorescent characteristics, such as fluorescent intensity and fluorescent wavelength, are varied by the amount of nucleic acids (such as DNA) in the reaction solution. In specific terms, this can be a fluorescent dye whose characteristics, such as fluorescent intensity and fluorescent wavelength, are varied by intercalation with double-stranded DNA. From the standpoint of ease of measurement, a fluorescent dye having the property of increasing in fluorescent intensity is preferable. Specific examples of such fluorescent dyes include ethidium bromide (EtBr), SYBR Green I, Pico Green, thiazole orange, and oxazole yellow. For example, ethidium bromide intercalated with DNA emits fluorescent light when excited by energy conversion of UV rays (260 nm) absorbed by the DNA, or by its own absorbed light. SYBR Green I that has been intercalated with DNA emits green fluorescent light when excited by visible light around 470 nm or by UV rays around 260 nm. The fluorescent intensity of the light emitted by these fluorescent dyes is proportional to the amount of double-stranded DNA, so the progress of the PCR in the reaction chamber (such as whether or not the target nucleic acids have been amplified, or the amount of PCR amplification product) can be monitored in real time (that is, instantly during the course of the PCR) by measuring the fluorescent intensity of the fluorescent dye.  
      The fluorescent material that can serve as an index of the progress of a PCR can also be one in which two types of fluorescent dye (a reporter and a quencher) are bonded to an oligonucleotide probe that is complementary with the middle portion of the target sequence. A reporter is a molecule that emits fluorescent light upon being irradiated with excitation light, but in the case of an oligonucleotide probe in which a quencher is present in the vicinity of a reporter, the energy absorbed by the reporter is absorbed by the quencher, the reporter is not excited, and the fluorescent light that was supposed to be produced is not produced (quenching). If an oligonucleotide probe undergoing quenching is added to a PCR reaction solution held in the reaction chamber, it bonds to the target sequence. A chain is then extended from the 3′-end of the primer by means of Taq polymerase, but if it hits the probe during this time, the probe, which has already been annealed by 5′→3′ endonuclease activity, is decomposed, the adjacent reporter and quencher separate, and the reporter, which had been suppressed by the quencher, then begins to emit fluorescent light. Since this reaction occurs substantially in proportion to the PCR cycle, the progress of the PCR in the reaction chamber can be monitored in real time by measuring the fluorescent intensity of the reporter.  
      The fluorescent material that can serve as an index of the progress of a PCR can also be one in which a fluorescent dye is bonded to two types of oligonucleotide probe that hybridize adjacent to the target nucleic acid. If donor dye is bonded to the 3′-end of the probe on the 5′-side, while an acceptor dye is bonded to the 5′-end of the probe on the 3′-side, and if two types of probe hybridize adjacent to the target nucleic acid, the donor dye emits fluorescent light upon irradiation with the excitation light from an external light source, this light is absorbed by the acceptor dye, and the acceptor dye at this point gives off light of a different wavelength. As the PCR amplification product increases, the amount of probe hybridizing to the target nucleic acid also increases, so the progress of the PCR in the reaction chamber can be monitored in real time by measuring the fluorescent intensity.  
      (3) In a second aspect of the reaction vessel of the present invention, all or part of the contact surface of the cover member is flat.  
      With the reaction vessel pertaining to this aspect, the irradiation of the reaction solution with the excitation light and the detection of the fluorescent light emitted from the reaction solution can be performed through all or part of the flat contact surface of the cover member, which means that excitation light irradiation conditions and fluorescent light reception conditions can be set more easily.  
      (4) In a third aspect of the reaction vessel of the present invention, the contact surface of the cover member is the surface of the wall component of substantially uniform thickness that constitutes the cover member.  
      With the reaction vessel pertaining to this aspect, temperature control of the reaction solution held in the reaction chamber can be performed through the wall component (in the form of a plate, for example) of substantially uniform thickness having the contact surface of the cover member, and as a result the temperature of the reaction solution can be controlled rapidly and efficiently. The temperature control conditions here can also be set easily. Also, the irradiation of the reaction solution held in the reaction chamber with the excitation light and the detection of fluorescent light from the reaction solution can be performed through the wall component of substantially uniform thickness having the contact surface of the cover member, which means that excitation light irradiation conditions and fluorescent light reception conditions can be set more easily. Particularly when all or part of the contact surface of the cover member is flat, this wall component is also flat, so the contact surface of the cover member at this wall component is substantially parallel to the opposite surface, which means that excitation light irradiation conditions and fluorescent light reception conditions can be set even more easily.  
      (5) In a fourth aspect of the reaction vessel of the present invention, all or part of the contact surface of the reaction chamber is flat.  
      With the reaction vessel pertaining to this aspect, the irradiation of the reaction solution with the excitation light and the detection of the fluorescent light emitted from the reaction solution can be performed through all or part of the flat contact surface of the reaction chamber, which means that excitation light irradiation conditions and fluorescent light reception conditions can be set more easily.  
      (6) In a fifth aspect of the reaction vessel of the present invention, the contact surface of the reaction chamber is the surface of the wall component of substantially uniform thickness that constitutes the reaction vessel main body.  
      With the reaction vessel pertaining to this aspect, the temperature of the reaction solution held in the reaction chamber can be controlled through the wall component (in the form of a plate, for example) of substantially uniform thickness having the contact surface of the reaction chamber, allowing the temperature of the reaction solution to be controlled more rapidly and efficiently. Also, temperature control here conditions can be set more easily. Also, the irradiation of the reaction solution held in the reaction chamber with the excitation light and the detection of the fluorescent light from the reaction solution can be performed through the wall component of substantially uniform thickness having the contact surface of the reaction chamber, which means that excitation light irradiation conditions and fluorescent light reception conditions can be set more easily. Particularly when all or part of the contact surface of the reaction chamber is flat, this wall component will be in the form of a flat plate, so the contact surface of the reaction chamber at this wall component is substantially parallel to the opposite surface, which means that excitation light irradiation conditions and fluorescent light reception conditions can be set even more easily.  
      (7) In a sixth aspect of the reaction vessel of the present invention, a tightly closed space is formed by the contact surface of the reaction chamber and the contact surface of the cover member when the cover member is mounted on the reaction vessel main body, and all or part of the reaction solution is held in the tightly closed space.  
      With the reaction vessel pertaining to this aspect, when the cover member is mounted on the reaction vessel main body, the end of the contact surface of the cover member fits snugly with the end of the contact surface of the reaction chamber, forming a tightly closed space. Whether all or just part of the reaction solution will be held in this tightly closed space is determined according to the volume of reaction solution held in the reaction chamber, the volume of the tightly closed space that is formed, and so forth.  
      With the reaction vessel pertaining to this aspect, all (or nearly all) of the outer surface of the reaction solution held in the tightly closed space becomes the contact surface with the cover member and the reaction chamber, so the temperature of the reaction solution can be rapidly controlled through the contact surface of the cover member and/or the contact surface of the reaction chamber.  
      Also, since there is no air or other such gas (or almost none) present in the tightly closed space, evaporation of the reaction solution into gas can be suppressed while the desired reaction proceeds within the tightly closed space, which means that the reaction will proceed even if there is only a tiny amount of reaction solution held in the reaction chamber.  
      (8) In a seventh aspect of the reaction vessel of the present invention, a surplus reaction solution holder capable of holding any surplus reaction solution that cannot be held in the tightly closed space is formed in the reaction chamber when the cover member is mounted on the reaction vessel main body.  
      With the reaction vessel pertaining to this aspect, reaction solution whose volume is greater than that which can be held in the tightly closed space is held in the reaction chamber, and the reaction solution is pressed by the cover member (such as a convex component provided to the cover member), the result of which is that any air in the reaction chamber, bubbles in the reaction solution, or the like is pushed out into the surplus reaction solution holder along with the reaction solution that cannot be held in the tightly closed space, and this prevents air or bubbles from getting into the reaction solution held in the tightly closed space. Also, since a constant amount of reaction solution is held in the tightly closed space, the reaction will proceed for a constant amount of reaction solution regardless of the volume of reaction solution held in the reaction chamber, which reduces the labor for metering the reaction solution precisely and adding it to the reaction chamber.  
      With the reaction vessel pertaining to this aspect, the surplus reaction solution holder is formed in the reaction chamber as follows. When the cover member has been placed over the reaction vessel main body, the outer peripheral surface of the convex component of the cover member does not fit snugly against the inner peripheral surface of the reaction chamber, forming a space between the outer peripheral surface of the convex component of the cover member and the inner peripheral surface of the reaction chamber, and this space serves as the surplus reaction solution holder.  
      (9) In an eighth aspect of the reaction vessel of the present invention, the reaction chamber has an opposing surface opposite the contact surface of the cover member, and when the cover member is mounted on the reaction vessel main body, all or part of the reaction solution held in the reaction chamber is held in the form of a thin layer between the contact surface of the cover member and the opposing surface of the reaction chamber.  
      In the present invention, of the contact surface of the reaction chamber, the surface that is opposite the contact surface of the cover member is called the “opposing surface of the reaction chamber.” If the contact surface of the cover member is flat, then the opposing surface of the reaction chamber is preferably flat so as to correspond, and if the contact surface of the cover member is curved, then the opposing surface is preferably curved so as to correspond.  
      With the reaction vessel pertaining to this aspect, the reaction is monitored for the reaction solution held in the form of a thin layer between the contact surface of the cover member and the opposing surface of the reaction chamber. Here, the reaction solution in the form of a thin layer has a large ratio of surface area to volume, so the temperature of the reaction solution can be rapidly controlled by the movement of heat through the contact surface of the reaction chamber and/or the contact surface of the cover member. Also, having the reaction solution in the form of a thin layer allows temperature control to be performed more uniformly for the entire reaction solution.  
      (10) In a ninth aspect of the reaction vessel of the present invention, the opposing surface of the reaction chamber is the surface of the wall component of substantially uniform thickness that constitutes the reaction vessel main body.  
      With the reaction vessel pertaining to this aspect, the temperature of the reaction solution held in the reaction chamber can be controlled through the wall component (in the form of a plate, for example) of substantially uniform thickness having the opposing surface of the reaction chamber, which allows the temperature of the reaction solution to be controlled more rapidly and efficiently. The setting of the temperature control conditions here is also easier. Furthermore, the irradiation of the reaction solution held in the reaction chamber with the excitation light and the detection of the fluorescent light from the reaction solution can be performed through the wall component of substantially uniform thickness having the opposing surface of the reaction chamber, which means that excitation light irradiation conditions and fluorescent light reception conditions can be set more easily. In particular, if all or part of the opposing surface of the reaction chamber is flat, this wall component will be in the form of a flat plate, and the opposing surface of the reaction chamber at this wall component will be substantially parallel to the surface on the opposite side, which means that excitation light irradiation conditions and fluorescent light reception conditions can be set even more easily.  
      (11) In a tenth aspect of the reaction vessel of the present invention, the wall component having the opposing surface of the reaction chamber is made of a light transmitting material so that light can pass from the outside of the reaction vessel to the reaction solution held in the reaction chamber, and/or from the reaction solution held in the reaction chamber to the outside of the reaction vessel, through the opposing surface of the reaction chamber.  
      With the reaction vessel pertaining to this aspect, if light can pass from the outside of the reaction vessel to the reaction solution held in the reaction chamber through the opposing surface of the reaction chamber, then the reaction solution held in the reaction chamber is irradiated with light from the outside of the reaction vessel through the opposing surface of the reaction chamber. If light can pass from the reaction solution held in the reaction chamber to the outside of the reaction vessel through the opposing surface of the reaction chamber, then the light (such as fluorescent light or chemical luminescence) emitted from the reaction solution held in the reaction chamber can be detected on the outside of the reaction vessel through the opposing surface of the reaction chamber. Therefore, the irradiation of the reaction solution with light and/or the detection of light emitted from the reaction solution performed in the monitoring of the reaction can be performed through the opposing surface of the reaction chamber.  
      (12) In an eleventh aspect of the reaction vessel of the present invention, the reaction vessel main body has an abutting surface that defines the distance between the contact surface of the cover member and the opposing surface of the reaction chamber by abutting against the cover member.  
      With the reaction vessel pertaining to this aspect, the thickness of the reaction solution held between the contact surface of the cover member and the opposing surface of the reaction chamber is kept constant by keeping constant the distance between the contact surface of the cover member and the opposing surface of the reaction chamber, so temperature control can be carried out uniformly for the entire reaction solution. Also, the thickness of the reaction solution can be adjusted by adjusting the distance between the contact surface of the cover member and the opposing surface of the reaction chamber.  
      With the reaction vessel pertaining to this aspect, the reaction vessel main body may have the abutting surface either inside or outside the reaction chamber. For example, the abutting surface may be provided along the inner peripheral surface of the reaction chamber. In this case, when the cover member is mounted on the reaction vessel main body, the cover member fits snugly around the inner peripheral surface of the reaction chamber, sealing off the inside of the reaction chamber.  
      (13) In a twelfth aspect of the reaction vessel of the present invention, the reaction chamber has an enveloping surface that envelops the reaction solution present between the contact surface of the cover member and the opposing surface of the reaction chamber, and when the cover member is mounted on the reaction vessel main body, a tightly closed space is formed by the contact surface of the cover member, the opposing surface of the reaction chamber, and the enveloping surface of the reaction chamber, and all or part of the reaction solution is held in the form of a thin layer within the tightly closed space.  
      In the present invention, of the contact surface of the reaction chamber, the surface that envelops the reaction solution present between the contact surface of the cover member and the opposing surface of the reaction chamber is called the “enveloping surface of the reaction chamber.” The shape of the enveloping surface of the reaction chamber will be determined by the shape of the contact surface of the cover member and the opposing surface of the reaction chamber. For instance, if the contact surface of the cover member and the opposing surface of the reaction chamber are circular, then the enveloping surface of the reaction chamber will be cylindrical, and if the contact surface of the cover member and the opposing surface of the reaction chamber are rectangular, the enveloping surface of the reaction chamber will be in the form of an angular cylinder. The lateral cross sectional shape of the enveloping surface can be selected as desired, but examples include circular, quadrangular (including both square and rectangular), semicircular, and parallelogram-shaped.  
      With the reaction vessel pertaining to this aspect, the end of the opposing surface of the reaction chamber communicates with the end (usually the lower end) of the enveloping surface of the reaction chamber, and when the cover member is mounted on the reaction vessel main body, the end of the contact surface of the cover member fits snugly with the end (usually the upper end) of the enveloping surface of the reaction chamber. As a result, a tightly closed space is formed by the contact surface of the cover member, the opposing surface of the reaction chamber, and the enveloping surface of the reaction chamber.  
      With the reaction vessel pertaining to this aspect, the reaction is monitored for the reaction solution held in the form of a thin layer in the tightly closed space. Here, the reaction solution in the form of a thin layer has a large ratio of surface area to volume, so the temperature of the reaction solution can be rapidly controlled by the movement of heat through the contact surface of the reaction chamber and/or the contact surface of the cover member. Also, if the reaction solution is in the form of a thin layer, the thickness of the reaction solution will be substantially uniform, which allows temperature control to be performed more uniformly for the entire reaction solution. Furthermore, since there is no air or other such gas (or almost none) present in the tightly closed space, evaporation of the reaction solution into gas can be suppressed while the desired reaction proceeds within the tightly closed space, which means that the reaction will proceed even if there is only a tiny amount of reaction solution held in the reaction chamber.  
      (14) In a thirteenth aspect of the reaction vessel of the present invention, all or part of the enveloping surface of the reaction chamber is flat.  
      With the reaction vessel pertaining to this aspect, the irradiation of the reaction solution with excitation light or the detection of fluorescent light from the reaction solution can be performed through all or part of the flat enveloping surface of the reaction chamber, which means that the excitation light irradiation conditions and fluorescent light reception conditions can be set more easily.  
      (15) In a fourteenth aspect of the reaction vessel of the present invention, a lateral cross section of the enveloping surface of the reaction chamber is quadrangular.  
      With the reaction vessel pertaining to this aspect, the enveloping surface of the reaction chamber comprises four planes, and the opposing pairs of planes are parallel. Therefore, by utilizing the rectilinear propagation property of light, it is possible to irradiate the entire reaction solution with light and to detect the light emitted from the entire reaction solution through a single plane constituting the enveloping surface of the reaction chamber. Also, the excitation light irradiation conditions and fluorescent light reception conditions can be set more easily.  
      (16) In a fifteenth aspect of the reaction vessel of the present invention, the enveloping surface of the reaction chamber is the surface of the wall component of substantially uniform thickness that constitutes the reaction vessel main body.  
      With the reaction vessel pertaining to this aspect, the temperature of the reaction solution held in the reaction chamber can be controlled through the wall component (in the form of a plate, for example) of substantially uniform thickness having the enveloping surface of the reaction chamber, which allows the temperature of the reaction solution to be controlled rapidly and efficiently. The temperature control conditions here can also be set more easily. Also, the irradiation of the reaction solution held in the reaction chamber with excitation light or the detection of fluorescent light from the reaction solution can be performed through the wall component of substantially uniform thickness having the enveloping surface of the reaction chamber, which means that excitation light irradiation conditions and fluorescent light reception conditions can be set more easily. In particular, if all or part of the enveloping surface of the reaction chamber is flat, this wall component will be in the form of a flat plate, and the enveloping surface of the reaction chamber at this wall component will be substantially parallel to the surface on the opposite side, which means that excitation light irradiation conditions and fluorescent light reception conditions can be set even more easily.  
      (17) In a sixteenth aspect of the reaction vessel of the present invention, the wall component having the enveloping surface of the reaction chamber is made of a light transmitting material so that light can pass from the outside of the reaction vessel to the reaction solution held in the reaction chamber, and/or from the reaction solution held in the reaction chamber to the outside of the reaction vessel, through the enveloping surface of the reaction chamber.  
      With the reaction vessel pertaining to this aspect, if light can pass from the outside of the reaction vessel to the reaction solution held in the reaction chamber through the enveloping surface of the reaction chamber, then the reaction solution held in the reaction chamber can be irradiated with light from the outside of the reaction vessel through the enveloping surface of the reaction chamber. If light can pass from the reaction solution held in the reaction chamber to the outside of the reaction vessel through the enveloping surface of the reaction chamber, then the light (such as fluorescent light or chemical luminescence) emitted from the reaction solution held in the reaction chamber can be detected on the outside of the reaction vessel through the enveloping surface of the reaction chamber. Therefore, the irradiation of the reaction solution with light and/or the detection of light emitted from the reaction solution performed in the monitoring of the reaction can be performed through the enveloping surface of the reaction chamber.  
      With the reaction vessel pertaining to this aspect, the temperature of the reaction solution held in the reaction chamber is controlled by the movement of heat through the contact surface of the cover member and/or the opposing surface of the reaction chamber, and the irradiation of the reaction solution with light and/or the detection of light emitted from the reaction solution performed in the monitoring of the reaction can be performed through the enveloping surface of the reaction chamber. Thus having the surface utilized for controlling the temperature of the reaction solution be separate from the surface utilized for monitoring the progress of the reaction allows the temperature of the reaction solution to be controlled rapidly and also allows the region where the reaction progress is monitored to be set freely. It is also possible to monitor the reaction progress for the entire reaction solution.  
      (18) In a seventeenth aspect of the reaction vessel of the present invention, a nozzle tip fitting space, into which a nozzle tip mounted on a nozzle capable of the intake and discharge of a liquid can be fitted, is formed in the cover member of the reaction vessel pertaining to the sixth aspect, and a nozzle tip fitting hole leading to the nozzle tip fitting space is formed so as to allow the nozzle tip to be fitted into the nozzle tip fitting space while the cover member is mounted on the reaction vessel main body, and a through-hole communicating between the outside of the reaction vessel, the tightly closed space, and the nozzle tip fitting space can be formed in the reaction vessel main body and the cover member by a puncture needle provided on the outside of the reaction vessel while the cover member is mounted on the reaction vessel main body.  
      When the desired reaction is conducted utilizing the reaction vessel pertaining to this aspect, all or part of the reaction solution is held and the desired reaction is brought about in the tightly closed space formed by the contact surface of the cover member and the contact surface of the reaction chamber.  
      Either before or after the reaction, the nozzle tip mounted on the nozzle is mounted through the nozzle tip fitting hole in the nozzle tip fitting space of the reaction vessel while the cover member is mounted on the reaction vessel main body. The nozzle tip mounted in the nozzle tip fitting space is an intermediary member capable of transmitting the intake force (reduced pressure) or discharge force (pressurization) of the nozzle to the outside of the nozzle tip. One example of a nozzle tip that can be used is one in which a nozzle mounting hole is formed at one end, and an intake and discharge port leading to the nozzle mounting hole is formed at the other end. When a nozzle tip such as this is used, for example, the nozzle tip is mounted in the nozzle tip fitting space so that the intake and discharge port of the nozzle tip leads to the nozzle tip fitting space. Here, the location of the intake and discharge port of the nozzle tip inside the nozzle tip fitting space is defined by the abutting of the cover member against the abutting component of the nozzle tip, for example.  
      Either before or after the mounting of the nozzle tip in the nozzle tip fitting space, a through-hole communicating between the outside of the reaction vessel, the tightly closed space in which the reaction solution is held, and the nozzle tip fitting space can be formed in the reaction vessel main body and the cover member by a puncture needle provided on the outside of the reaction vessel. This puncture needle first punctures the reaction vessel main body, forming a through-hole communicating between the outside of the reaction vessel and the tightly closed space in which the reaction solution is held, and then punctures the cover member, forming a through-hole communicating between the tightly closed space in which the reaction solution is held and the nozzle tip fitting space. When the reaction vessel has thus undergone mounting of the nozzle tip and puncture by the puncture needle, the outside of the reaction vessel communicates with the tightly closed space in which the reaction solution is held, through the through-hole formed in the reaction vessel main body, the tightly closed space in which the reaction solution is held communicates with the nozzle tip fitting space through the through-hole formed in the cover member, and the nozzle tip fitting space leads to the intake and discharge portion of the nozzle tip, so the intake force and discharge force of the nozzle can be transmitted to the outside of the reaction vessel. Therefore, when the reaction vessel is immersed in a liquid so that the through-hole formed in the reaction vessel main body is also immersed in this liquid, and the intake and discharge of the nozzle are commenced, the above-mentioned liquid flows into the tightly closed space holding the reaction solution along with the intake of the nozzle, and flows out of the tightly closed space along with the discharge of the nozzle. When this intake and discharge of the nozzle are repeated over and over, the reaction solution held in the tightly closed space of the reaction vessel is extracted into the above-mentioned liquid. As the reaction solution is extracted, the reaction product contained in the reaction solution is also extracted into the above-mentioned liquid.  
      Thus, if the reaction vessel pertaining to this aspect is utilized, the reaction product contained in the reaction solution can be acquired without removing the cover member from the reaction vessel main body after the reaction has been conducted with the cover member covering the reaction vessel main body.  
      The nozzle tip may also be mounted in the nozzle tip fitting space so that the intake and discharge port of the nozzle tip is sealed off by contact with the wall component of the cover member forming the nozzle tip fitting space. However, the wall component of the cover member that seals off the intake and discharge port of the nozzle tip must have a contact surface with the tightly closed space in which the reaction solution is held. In this case, a through-hole that communicates with the outside of the reaction chamber, the tightly closed space in which the reaction solution is held, and the intake and discharge port of the nozzle tip is formed in the reaction vessel main body and the cover member by the puncture needle provided on the outside of the reaction vessel, either before or after the mounting of the nozzle tip in the nozzle tip fitting space. The through-hole that communicates with the tightly closed space in which the reaction solution is held and the intake and discharge port of the nozzle tip is formed in the wall component of the cover member that seals off the intake and discharge port of the nozzle tip. When the reaction vessel has thus undergone mounting of the nozzle tip and puncture by the puncture needle, the outside of the reaction vessel communicates with the tightly closed space in which the reaction solution is held, through the through-hole formed in the reaction vessel main body, and the tightly closed space in which the reaction solution is held communicates with the intake and discharge port of the nozzle tip through the through-hole formed in the cover member, so the intake force and discharge force of the nozzle can be transmitted to the outside of the reaction vessel. Therefore, just as above, when the reaction vessel is immersed in a liquid so that the through-hole formed in the reaction vessel main body is also immersed in this liquid, and the intake and discharge of the nozzle are repeated over and over, the reaction product contained in the reaction solution is extracted into the above-mentioned liquid.  
      With the reaction vessel pertaining to this aspect, the size and shape of the nozzle tip fitting space formed in the cover member, and of the nozzle tip fitting hole leading to this nozzle tip fitting space are suitably adjusted according to the size and shape of the nozzle to be mounted in the nozzle tip fitting space. Also, the nozzle tip fitting hole is formed at a location where the nozzle tip can be mounted in the nozzle tip fitting space through the nozzle tip fitting hole while the cover member is mounted on the reaction vessel main body.  
      With the reaction vessel pertaining to this aspect, the reaction vessel main body and the cover member are made of a material that will not be corroded by the reaction solution and that can withstand the reaction conditions (such as the reaction temperature). The selected material must be one that allows the reaction vessel main body and the cover member to be punctured by the puncture needle provided to the outside of the reaction vessel. If the puncture needle is made from a metal such as stainless steel, then a plastic or glass can be selected, for example, as the material for the reaction vessel main body and the cover member.  
      With the reaction vessel pertaining to this aspect, the relative positions of the tightly closed space holding the reaction solution and the nozzle tip fitting space are adjusted so that the through-hole that communicates between the outside of the reaction vessel, the tightly closed space in which the reaction solution is held, and the nozzle tip fitting space can be formed in the reaction vessel main body and the cover member by the puncture needle provided on the outside of the reaction vessel.  
      There are no particular restrictions on the shape of the puncture needle provided on the outside of the reaction vessel pertaining to this aspect, as long as it allows the cover member and the reaction vessel main body to be punctured, but an example of the shape of the puncture needle is one that is tapered at the distal end. Specific examples include conical, pyramidal, and acicular shapes. The term “tapered” as used here means a shape that narrows toward the distal end, and in addition to a shape in which the distal end is pointed, also includes a shape in which the distal end is rounded, a shape in which the distal end is flat, and so forth. The material of the puncture needle is suitably determined according to the materials of the cover member and the reaction vessel main body so that these can be punctured, but is usually stainless steel or another metal. There are no particular restrictions on the number of puncture needles used for puncturing. The length of the puncture needle is suitably adjusted so that the desired through-hole can be formed.  
      (19) In an eighteenth aspect of the reaction vessel of the present invention, a nozzle tip fitting space, into which a nozzle tip mounted on a nozzle capable of the intake and discharge of a liquid can be fitted, is formed in the cover member of the reaction vessel pertaining to the twelfth aspect, and a nozzle tip fitting hole leading to the nozzle tip fitting space is formed so as to allow the nozzle tip to be fitted into the nozzle tip fitting space while the cover member is mounted on the reaction vessel main body, and a through-hole communicating between the outside of the reaction vessel, the tightly closed space, and the nozzle tip fitting space can be formed in the reaction vessel main body and the cover member by a puncture needle provided on the outside of the reaction vessel while the cover member is mounted on the reaction vessel main body.  
      When the desired reaction is conducted utilizing the reaction vessel pertaining to this aspect, all or part of the reaction solution is held and the desired reaction is brought about in the tightly closed space formed by the contact surface of the cover member, the opposing surface of the reaction chamber, and the enveloping surface of the reaction chamber.  
      When the reaction vessel pertaining to this aspect is utilized, just as with the reaction vessel pertaining to the seventeenth aspect, the reaction product contained in the reaction solution can be acquired without removing the cover member from the reaction vessel main body after the reaction has been conducted with the cover member covering the reaction vessel main body.  
      (20) In a nineteenth aspect of the reaction vessel of the present invention, the nozzle tip fitting space is formed so that the nozzle tip fitting space is closed off when the nozzle tip fitting hole is sealed.  
      With the reaction vessel pertaining to this aspect, the nozzle tip fitting space is closed off by mounting the nozzle tip in the nozzle tip fitting space, so the intake and discharge forces of the nozzle can be efficiently transmitted to the nozzle tip fitting space. The term “closed off” as used here means that there are no gaps, slits, or the like that would hinder the transmission of the intake force (reduced pressure) or discharge force (pressurization) of the nozzle to the nozzle tip fitting space, and a state in which the nozzle tip fitting space leads to the intake and discharge port of the nozzle tip is encompassed by “closed off.” A state in which gaps, slits, and so forth are present to the extent that they pose no hindrance to the transmission of the intake force (reduced pressure) or discharge force (pressurization) of the nozzle to the nozzle tip fitting space is also encompassed by “closed off.” 
      (21) In a twentieth aspect of the reaction vessel of the present invention, the wall component of the cover member forming the nozzle tip fitting space has an inner peripheral surface capable of fitting snugly against the outer peripheral surface of the nozzle tip.  
      With the reaction vessel pertaining to this aspect, when the nozzle tip is mounted in the nozzle tip fitting space, the outer peripheral surface of the nozzle tip fits snugly against the inner peripheral surface of the wall component of the cover member forming the nozzle tip fitting space, thereby closing off the nozzle tip fitting space.  
      (22) In a twenty-first aspect of the reaction vessel of the present invention, a convex component and/or a concave component capable of fitting with a concave component and/or a convex component provided on the outer peripheral surface of the nozzle tip is provided on the inner peripheral surface of the wall component of the cover member capable of snugly fitting against the outer peripheral surface of the nozzle tip.  
      With the reaction vessel pertaining to this aspect, the nozzle tip is mounted more securely in the nozzle tip fitting space, so even if a force in the direction opposite the mounting direction into the nozzle tip fitting space is exerted on the nozzle tip mounted in the nozzle tip fitting space, the nozzle tip will not come out of the nozzle tip fitting space. Therefore, it is possible to move the reaction vessel while the cover member is mounted on the reaction vessel main body, by moving the nozzle on which is mounted the nozzle tip in the nozzle tip fitting space.  
      (23) In a twenty-second aspect of the reaction vessel of the present invention, the contact surface of the cover member is the surface of the wall component of the cover member forming the nozzle tip fitting space.  
      With the reaction vessel pertaining to this aspect, a through-hole communicating between the tightly closed space in which the reaction solution is held and the nozzle tip fitting space is formed in the wall component of the cover member having a contact surface with the reaction solution, which is the wall component of the cover member forming the nozzle tip fitting space. Also, since the wall component of the cover member is opposite some portion of the wall component of the reaction vessel main body forming the tightly closed space in which the reaction solution is held, a through-hole communicating between the outside of the reaction vessel, the tightly closed space in which the reaction solution is held, and the nozzle tip fitting space can be formed in the reaction vessel main body and the cover member by a single puncture needle provided on the outside of the reaction vessel.  
      (24) In a twenty-third aspect of the reaction vessel of the present invention, the contact surface of the cover member is the surface of the wall component of the cover member forming the deepest portion of the nozzle tip fitting space.  
      Here, “the deepest portion of the nozzle tip fitting space” means the portion of the nozzle tip fitting space that is farthest away from the nozzle tip fitting hole. With the reaction vessel pertaining to this aspect, the nozzle tip is mounted toward the deepest part of the nozzle tip fitting space from the nozzle tip fitting hole.  
      (25) In a twenty-fourth aspect of the reaction vessel of the present invention, the wall component of the cover member forming the deepest portion of the nozzle tip fitting space is provided so as to oppose the wall component of the reaction vessel main body forming the deepest part of the tightly closed space.  
      Here, “the deepest part of the tightly closed space” means the portion of the tightly closed space in which the reaction solution is held that is closest to the surface on which the reaction vessel is placed. With the reaction vessel pertaining to this aspect, a through-hole communicating between the outside of the reaction vessel, the tightly closed space in which the reaction solution is held, and the nozzle tip fitting space can be formed in the reaction vessel main body and the cover member by a puncture needle provided perpendicular or substantially perpendicular to the surface on which the reaction vessel is placed.  
      (26) In a twenty-fifth aspect of the reaction vessel of the present invention, the nozzle tip fitting space is formed such that the mounting direction of the nozzle tip with respect to the nozzle tip fitting space is perpendicular or substantially perpendicular to the surface on which the reaction vessel is placed.  
      With the reaction vessel pertaining to this aspect, the force exerted on the reaction vessel by the nozzle tip in the mounting of the nozzle tip in the nozzle tip fitting space is a force that is perpendicular or substantially perpendicular to the surface on which the reaction vessel is placed. Therefore, the nozzle tip can be easily mounted in the nozzle tip fitting space without the reaction vessel shifting its position while the nozzle tip is being mounted in the nozzle tip fitting space.  
      (27) In a twenty-sixth aspect of the reaction vessel of the present invention, the cover member has an outer peripheral surface capable of fitting snugly against the inner peripheral surface of the reaction chamber.  
      With the reaction vessel pertaining to this aspect, the outer peripheral surface of the cover member fits snugly against the inner peripheral surface of the reaction chamber, the result of which is that the tightly closed space in which the reaction solution is held is closed off more effectively. Therefore, the intake and discharge force of the nozzle can be transmitted more efficiently to the outside of the reaction vessel.  
      (28) In a twenty-seventh aspect of the reaction vessel of the present invention, a concave component and/or a convex component is provided on the inner peripheral surface of the reaction chamber, and a convex component and/or a concave component capable of mating with the concave component and/or the convex component provided on the inner peripheral surface of the reaction chamber is provided on the outer peripheral surface of the cover member.  
      With the reaction vessel pertaining to this aspect, the cover member covers the reaction vessel main body more securely, so even if the reaction vessel is moved while the cover member is mounted on the reaction vessel main body (such as when the cover member is supported, but not the reaction vessel main body, while the reaction vessel is moved), the cover member will not come off the reaction vessel main body. Therefore, it is possible to move the reaction vessel while the cover member is mounted on the reaction vessel main body, by moving the nozzle on which is mounted the nozzle tip in the nozzle tip fitting space.  
      (29) In a twenty-eighth aspect of the reaction vessel of the present invention, the reaction vessel is a reaction vessel for PCR.  
      With the reaction vessel pertaining to this aspect, the reaction occurring in the reaction chamber is a PCR, and the reaction solution held in the reaction chamber is a reaction solution for PCR. PCR reaction solutions include H 2 O, buffers, MgCl 2 , dNTP mixes, primers, template DNA, Taq polymerase, and so forth, and PCR amplified fragments (such as DNA fragments) are contained as the reaction product in the PCR reaction solution after the reaction.  
      With PCR, the temperature of the reaction solution must be controlled over time or periodically, and since the temperature of a reaction solution can be controlled rapidly with the reaction vessel of the present invention, the time required by a PCR can be reduced by using the reaction vessel of the present invention as a PCR reaction vessel. Also, PCR is a technique involving the amplification of extremely small amounts of template DNA, so contamination with other DNA is a serious problem, but since contamination of the reaction solution is prevented with the reaction vessel of the present invention, the desired PCR can be performed accurately by using the reaction vessel of the present invention as a PCR reaction vessel. Furthermore, since evaporation of the reaction solution held in the reaction chamber is suppressed with the reaction vessel of the present invention, a PCR can be conducted with only a tiny amount of PCR reaction solution by using the reaction vessel of the present invention as a PCR reaction vessel. In addition, the progress of a PCR can be monitored in real time by using reaction vessel of the present invention as a PCR reaction vessel.  
      When the reaction vessel pertaining to this aspect is utilized, a series of operations comprising the preparation of samples containing target nucleic acids (such as extraction of nucleic acids from cells), amplification of these target nucleic acids by PCR, and monitoring (detection, measurement, qualitative analysis, quantitative analysis, etc.) of the progress of the PCR (such as whether or not the target nucleic acids have been amplified, or the amount of PCR amplification product) can be automated.  
      (30) In order to achieve the stated objects, the first reaction apparatus of the present invention is a reaction apparatus comprising the reaction vessel pertaining to the first aspect, a temperature controller, a light source, and a fluorescent light detector, wherein the temperature controller is attached to the cover member and/or the reaction vessel main body so that temperature of the reaction solution held in the reaction chamber can be controlled through the contact surface of the cover member and/or the contact surface of the reaction chamber, the light source is provided so that the reaction solution held in the reaction chamber can be irradiated with light through the contact surface of the cover member and/or the contact surface of the reaction chamber, and the fluorescent light detector is provided so that fluorescent light emitted from the reaction solution held in the reaction chamber can be detected through the contact surface of the cover member and/or the contact surface of the reaction chamber.  
      With the first reaction apparatus of the present invention, the temperature controller is attached to the cover member and/or the reaction vessel main body, and the temperature of the reaction solution can be rapidly controlled by the movement of heat through the contact surface of the cover member and/or the contact surface of the reaction chamber. The temperature controller may be attached directly to the cover member and/or the reaction vessel main body, or it may be attached via another member. For instance, with the reaction vessel of the present invention, if a heat-conducting metal block or heat-conducting metal plate is provided so as to be in contact with the reaction vessel main body and/or the cover member, the temperature controller can be attached to the cover member and/or the reaction vessel main body via the heat-conducting metal block or heat-conducting metal plate. With the first reaction apparatus of the present invention, the temperature controller may be attached to just the cover member or just the reaction vessel main body, or to both, but from the standpoint of rapidly controlling the temperature of the reaction solution, it is preferably attached to both the cover member and the reaction vessel main body.  
      With the first reaction apparatus of the present invention, the light source can irradiate the reaction solution held in the reaction chamber with excitation light through the contact surface of the cover member and/or the contact surface of the reaction chamber.  
      Also, with the first reaction apparatus of the present invention, the fluorescent light detector can detect fluorescent light emitted from the reaction solution held in the reaction chamber through the contact surface of the cover member and/or the contact surface of the reaction chamber.  
      With the first reaction apparatus of the present invention, the irradiation of the reaction solution with the excitation light and the detection of the fluorescent light emitted from the reaction solution can be performed by combining as desired the contact surface of the cover member and the contact surface of the reaction chamber. Specifically, the irradiation of the reaction solution with the excitation light and the detection of the fluorescent light emitted from the reaction solution can both be performed through the contact surface of the cover member, or both through the contact surface of the reaction chamber, or respectively through the contact surface of the cover member and the contact surface of the reaction chamber, or respectively through the contact surface of the reaction chamber and the contact surface of the cover member.  
      With the first reaction apparatus of the present invention, the reaction solution can be irradiated with excitation light and the fluorescent light emitted from the reaction solution can be detected while the reaction solution is still inside the reaction chamber. Therefore, if a fluorescent material that will serve as an index of the reaction progress is added ahead of time to the reaction solution, then the progress of the reaction can be monitored by detecting the light emitted by the fluorescent material during the course of the reaction. In particular, with the first reaction apparatus of the present invention, the temperature of the reaction solution held in the reaction chamber can be rapidly controlled and the progress of the reaction occurring in the reaction chamber can be monitored in real time (that is, instantly during the course of the reaction) while the reaction is proceeding.  
      (31) In a first aspect of the first reaction apparatus of the present invention, the temperature controller is attached to the wall component of substantially uniform thickness that constitutes the cover member and that has the contact surface of the cover member, and/or the wall component of substantially uniform thickness that constitutes the reaction vessel main body and that has the contact surface of the reaction chamber.  
      With the first reaction apparatus pertaining to this aspect, the temperature of the reaction solution held in the reaction chamber can be controlled through the wall component of substantially uniform thickness having the contact surface of the cover member, and/or the wall component of substantially uniform thickness having the contact surface of the reaction chamber, which means that the temperature of the reaction solution can be controlled rapidly and efficiently. The temperature control conditions here can also be set more easily.  
      Also, the irradiation of the reaction solution held in the reaction chamber with the excitation light and the detection of the fluorescent light from the reaction solution can be performed through the wall component of substantially uniform thickness having the contact surface of the cover member, and/or the wall component of substantially uniform thickness having the contact surface of the reaction chamber, which means that the excitation light irradiation conditions and fluorescent light reception conditions can be set more easily. In particular, if all or part of the contact surface of the cover member or the contact surface of the reaction chamber is flat, this wall component will be in the form of a flat plate, so the contact surface of the cover member or the contact surface of the reaction chamber at this wall component is substantially parallel to the opposite surface, which means that excitation light irradiation conditions and fluorescent light reception conditions can be set even more easily.  
      With the first reaction apparatus pertaining to this aspect, the temperature controller may be attached directly to the above-mentioned wall component, or it may be attached via another member. For instance, if a heat-conducting metal block or heat-conducting metal plate is provided so as to be in contact with the wall component, the temperature controller can be attached via the heat-conducting metal block or heat-conducting metal plate.  
      (32) In a second aspect of the first reaction apparatus of the present invention, the reaction vessel is the reaction vessel pertaining to the twelfth aspect, the temperature controller is attached to the cover member and/or the reaction vessel main body so that temperature of the reaction solution held in the reaction chamber can be controlled through the contact surface of the cover member and/or the opposing surface of the reaction chamber, the light source is provided so that the reaction solution held in the reaction chamber can be irradiated with light through the enveloping surface of the reaction chamber, and the fluorescent light detector is provided so that fluorescent light emitted from the reaction solution held in the reaction chamber can be detected through the enveloping surface of the reaction chamber.  
      With the first reaction apparatus pertaining to this aspect, the progress of the reaction occurring in the reaction chamber can be monitored in real time by irradiating the reaction solution with excitation light and detecting the fluorescent light emitted from the reaction solution through the enveloping surface of the reaction chamber while rapidly controlling the temperature of the reaction solution held in the reaction chamber by the movement of heat through the contact surface of the cover member and/or the opposing surface of the reaction chamber. In particular, with the first reaction apparatus pertaining to this aspect, since the surface utilized for controlling the temperature of the reaction solution (the contact surface of the cover member and/or the opposing surface of the reaction chamber) is separate from the surface utilized for monitoring the progress of the reaction (the enveloping surface of the reaction chamber), the region where the reaction progress is monitored can be set freely, and it is also possible to monitor the reaction progress for the entire reaction solution.  
      (33) In a third aspect of the first reaction apparatus of the present invention, the temperature controller is attached to the wall component of substantially uniform thickness that constitutes the cover member and that has the contact surface of the cover member, and/or the wall component of substantially uniform thickness that constitutes the reaction vessel main body and that has the opposing surface of the reaction chamber.  
      With the first reaction apparatus pertaining to this aspect, the temperature of the reaction solution held in the reaction chamber can be controlled through the wall component of substantially uniform thickness having the contact surface of the cover member, and/or the wall component of substantially uniform thickness having the opposing surface of the reaction chamber, which means that the temperature of the reaction solution can be controlled rapidly and efficiently. The temperature control conditions here can also be set easily.  
      Also, the irradiation of the reaction solution held in the reaction chamber with the excitation light and the detection of fluorescent light from the reaction solution can be performed through the wall component of substantially uniform thickness having the contact surface of the cover member and/or the wall component of substantially uniform thickness having the opposing surface of the reaction chamber, which means that the excitation light irradiation conditions and the fluorescent light reception conditions can be set more easily. Particularly when all or part of the contact surface of the cover member or the opposing surface of the reaction chamber is flat, this wall component is also flat, so the contact surface of the cover member at this wall component is substantially parallel to the opposite surface, or the opposing surface of the reaction chamber is substantially parallel to the opposite surface, which means that excitation light irradiation conditions and fluorescent light reception conditions can be set even more easily.  
      With the first reaction apparatus pertaining to this aspect, the temperature controller may be attached directly to the above-mentioned wall component, or it may be attached via another member. For instance, if a heat-conducting metal block or heat-conducting metal plate is provided so as to be in contact with the wall component, the temperature controller can be attached via the heat-conducting metal block or heat-conducting metal plate.  
      (34) In a fourth aspect of the first reaction apparatus of the present invention, the reaction apparatus further comprises a plurality of optical fibers disposed around the enveloping surface of the reaction chamber, wherein the irradiation of the reaction solution with light from the light source and/or the detection of fluorescent light emitted from the reaction solution is accomplished by utilizing the optical fibers.  
      With the first reaction apparatus pertaining to this aspect, the optical fibers are, for example, disposed around that surface of the wall component constituting the cover member and having the enveloping surface of the reaction chamber, that is opposite from the enveloping surface of the reaction chamber. If all or part of the enveloping surface of the reaction chamber is flat and the thickness of the wall component having the enveloping surface of the reaction chamber is substantially uniform, this wall component will be in the form of a flat plate, so the enveloping surface of the reaction chamber at this wall component is substantially parallel to the opposite surface, which means that excitation light irradiation conditions and fluorescent light reception conditions can be set even more easily if the optical fibers are disposed perpendicular to the wall component having the enveloping surface of the reaction chamber.  
      When a plurality of optical fibers are used to irradiate the reaction solution with light and to detect fluorescent light from the reaction solution, since the irradiation surface area of each optical fiber is small, the area excited by each optical fiber will also be small, and the intensity of the fluorescent light emitted from this area will be weak, if the distance the irradiation light is transmitted through the reaction solution is too short. Therefore, the detection sensitivity of each optical fiber will be low if the optical fibers are used in the irradiation of reaction solution present in the form of a thin layer between the contact surface of the cover member and the opposing surface of the reaction chamber with excitation light through the contact surface of the cover member and/or the opposing surface of the reaction chamber.  
      In contrast, if reaction solution present in the form of a thin layer between the contact surface of the cover member and the opposing surface of the reaction chamber is irradiated with excitation light through the enveloping surface of the reaction chamber, even though the irradiation surface area of each optical fiber is small, the distance the irradiation light travels through the reaction solution is longer, so the area excited by each optical fiber is larger and the intensity of fluorescent light emitted from that area is higher. Therefore, the detection sensitivity of the optical fibers is higher.  
      Accordingly, with the reaction vessel pertaining to this aspect, the progress of the reaction in a reaction solution present in the form of a thin layer between the contact surface of the cover member and the opposing surface of the reaction chamber can be monitored with good sensitivity by using optical fibers.  
      With the first reaction apparatus pertaining to this aspect, either the irradiation of the reaction solution with light from the light source or the detection of the fluorescent light emitted from the reaction solution, or both, may be performed with a single optical fiber. Also, the optical fibers utilized for irradiating the reaction solution with light from the light source and the optical fibers utilized for detecting the fluorescent light emitted from the reaction solution can be disposed as desired around the enveloping surface of the reaction chamber. Also, the type of irradiation light and the type of detected fluorescent light may be the same for all of the optical fibers, or may vary with each optical fiber or by optical fiber group.  
      With the first reaction apparatus pertaining to this aspect, if the reaction solution held in the reaction chamber is made ahead of time to contain a plurality of different fluorescent dyes, and irradiation with excitation light corresponding to each fluorescent dye and the detection of the fluorescent light emitted from each fluorescent dye are performed by individual optical fiber or by optical fiber group, then different reactions can be conducted at the same time, and the progress of the reactions (such as whether or not the target nucleic acids have been amplified, or the amount of PCR amplification product) can be monitored in real time. Also, if the same excitation light is emitted and the same fluorescent light detected by a plurality of optical fibers, then the progress of the reaction can be monitored for the entire reaction solution by disposing the optical fibers all the way around the enveloping surface of the reaction chamber.  
      (35) In order to achieve the stated objects, the second reaction apparatus of the present invention is a reaction apparatus comprising a reaction vessel installation part in which the reaction vessel pertaining to the seventeenth or eighteenth aspect, a first temperature controller, a second temperature controller, a light source, and a fluorescent light detector, wherein the first temperature controller is provided so that the temperature of the reaction solution held in the tightly closed space of the reaction vessel installed in the reaction vessel installation part can be controlled through the contact surface of the reaction chamber, the second temperature controller is removably mounted in the nozzle tip fitting space of the cover member and provided so that the temperature of the reaction solution held in the tightly closed space of the reaction vessel installed in the reaction vessel installation part can be controlled through the contact surface of the cover member, the light source is provided so that the reaction solution held in the tightly closed space of the reaction vessel installed in the reaction vessel installation part can be irradiated with light through the contact surface of the cover member and/or the contact surface of the reaction chamber, and the fluorescent light detector is provided so that fluorescent light emitted from the reaction solution held in the tightly closed space of the reaction vessel installed in the reaction vessel installation part can be detected through the contact surface of the cover member and/or the contact surface of the reaction chamber.  
      With the second reaction apparatus pertaining to this aspect, the reaction vessel before the reaction is placed in the reaction vessel installation part, and the temperature of the reaction solution held in the tightly closed space inside the reaction vessel is controlled by the first and second temperature controllers. The first and second temperature controllers are, for example, equipped with a heat-conducting metal block or heat-conducting metal plate provided so as to be in contact with the reaction vessel main body or the cover member, with the first temperature controller controlling the temperature of the reaction solution through the contact surface of the reaction chamber, and the second temperature controller controlling the temperature of the reaction solution through the contact surface of the cover member. The second temperature controller is designed so that it can be mounted in and removed from the nozzle tip fitting space of the cover member, and is mounted in the nozzle tip fitting space during the reaction, and removed from the nozzle tip fitting space after the reaction.  
      With the second reaction apparatus pertaining to this aspect, the reaction solution can be irradiated with excitation light from the light source through the contact surface of the cover member and/or the contact surface of the reaction chamber, and the fluorescent light emitted from the reaction solution can be detected by the fluorescent light detector through the contact surface of the cover member and/or the contact surface of the reaction chamber. As a result, the progress of the reaction occurring in the reaction solution can be monitored in real time (that is, instantly during the course of the reaction) while the desired reaction is being conducted by controlling the temperature of the reaction solution.  
      (36) In a first aspect of the second reaction apparatus of the present invention, the reaction vessel is the reaction vessel pertaining to the eighteenth aspect, the first temperature controller is provided so that the temperature of the reaction solution held in the tightly closed space of the reaction vessel installed in the reaction vessel installation part can be controlled through the opposing surface of the reaction chamber, the light source is provided so that the reaction solution held in the tightly closed space of the reaction vessel installed in the reaction vessel installation part can be irradiated with light through the enveloping surface of the reaction chamber, and the fluorescent light detector is provided so that fluorescent light emitted from the reaction solution held in the tightly closed space of the reaction vessel installed in the reaction vessel installation part can be detected through the enveloping surface of the reaction chamber.  
      With the second reaction apparatus pertaining to this aspect, the progress of the reaction occurring in the reaction chamber can be monitored in real time by irradiating the reaction solution with the excitation light and detecting the fluorescent light from the reaction solution through the enveloping surface of the reaction chamber while rapidly controlling the temperature of the reaction solution by the movement of heat through the contact surface of the cover member and the opposing surface of the reaction chamber. In particular, with the second reaction apparatus pertaining to this aspect, since the surface utilized for controlling the temperature of the reaction solution (the contact surface of the cover member and the opposing surface of the reaction chamber) is separate from the surface utilized for monitoring the progress of the reaction (the enveloping surface of the reaction chamber), the region where the reaction progress is monitored can be set freely, and it is also possible to monitor the reaction progress for the entire reaction solution.  
      (37) In a second aspect of the second reaction apparatus of the present invention, the reaction apparatus further comprises a plurality of optical fibers disposed around the enveloping surface of the reaction chamber, wherein the irradiation of the reaction solution with light from the light source and/or the detection of fluorescent light emitted from the reaction solution is accomplished by utilizing the optical fibers.  
      With the second reaction apparatus pertaining to this aspect, detection of the fluorescent light with the optical fibers can be carried out in the same manner as with the first reaction apparatus pertaining to the fourth aspect, and the same effect can be obtained as with the first reaction apparatus pertaining to the fourth aspect.  
      (38) In a third aspect of the second reaction apparatus of the present invention, the reaction apparatus further comprises a temperature controller mounting and removing part for mounting and removing the second temperature controller in the nozzle tip fitting space, wherein the temperature controller mounting and removing part performs an operation for mounting the second temperature controller in the nozzle tip fitting space prior to the reaction, and operation for removing the second temperature controller from the nozzle tip fitting space after the reaction.  
      (39) In a fourth aspect of the second reaction apparatus of the present invention, the reaction apparatus further comprises a puncture vessel installation part in which a puncture vessel is installed, a nozzle capable of the intake and discharge of a liquid, and a nozzle transfer part, wherein the puncture vessel comprises a liquid holding space capable of holding a liquid, an opening that leads to the liquid holding space, and a puncture needle, the liquid holding space is formed so that the reaction vessel can be accommodated in the liquid holding space through the opening, the puncture needle is provided so as to protrude into the liquid holding space from the wall component of the puncture vessel forming the liquid holding space, the nozzle transfer part performs an operation for fitting the nozzle tip mounted on the nozzle in the nozzle tip fitting space of the reaction vessel installed in the reaction vessel installation part, operation for transferring the reaction vessel with the mounted nozzle tip fitted thereinto to the puncture vessel installation part, and operation for accommodating the reaction vessel in the liquid holding space of the puncture vessel installed puncture vessel installation part, and for forming in the cover member and the reaction vessel main body, by means of the puncture needle provided in the puncture vessel, a through-hole that communicates with the nozzle tip fitting space, the tightly closed space of the reaction vessel, and the liquid holding space of the puncture vessel, and the nozzle performs an operation for extracting the reaction solution held in the tightly closed space of the reaction vessel into the liquid held in the liquid holding space of the puncture vessel, by the intake and discharge of the liquid through the through-hole.  
      With the second reaction apparatus pertaining to this aspect, the nozzle is moved by the nozzle transfer part, and the nozzle tip mounted on the nozzle is mounted in the nozzle tip fitting space of the reaction vessel placed in the reaction vessel installation part after the reaction. After the mounting of the nozzle tip in the nozzle tip fitting space, the nozzle is moved by the nozzle transfer part, and the reaction vessel is moved from the reaction vessel installation part to the puncture vessel installation part. The nozzle is then moved by the nozzle transfer part, and the reaction vessel is placed in the liquid holding space of the puncture vessel placed in the puncture vessel installation part. Here, the reaction vessel is pressed against the puncture needle provided to the puncture vessel, so that the puncture needle forms the desired through-hole (that is, a through-hole communicating between the liquid holding space of the puncture vessel, the tightly closed space in which the reaction solution is held, and the nozzle tip fitting space) in the cover member and the reaction vessel main body. The intake and discharge by the nozzle are then commenced, and the liquid held in the liquid holding space of the puncture vessel is taken in and discharged through the above-mentioned through-hole. The repeated intake and discharge by the nozzle cause the reaction solution held in the tightly closed space of the reaction vessel to be extracted into the above-mentioned liquid. Along with the extraction of the reaction solution, the reaction product contained in the reaction solution is also extracted into the liquid.  
      Thus, if the second reaction apparatus pertaining to this aspect is utilized, the reaction product contained in the reaction solution inside the reaction vessel can be acquired without removing the cover member from the reaction vessel main body after the reaction has been conducted while the reaction vessel main body is covered by the cover member.  
      With the second reaction apparatus pertaining to this aspect, the nozzle may have any structure that allows the liquid to be taken in and discharged, but an example of a nozzle that can be used is one having the same structure as the nozzle utilized in a conventional dispensing apparatus. The nozzle transfer part may also have any structure that allows the required operations to be carried out.  
      The operation for mounting the nozzle tip in the nozzle tip fitting space is performed after the operation for removing the second temperature controller from the nozzle tip fitting space. Also, the operation for mounting the nozzle tip in the nozzle tip fitting space is controlled so as not to interfere with the operation for removing the second temperature controller from the nozzle tip fitting space.  
      (40) In a fifth aspect of the first and second reaction apparatus of the present invention, the reaction apparatus is a reaction apparatus for PCR.  
      With the reaction apparatus pertaining to this aspect, the reaction occurring in the reaction chamber is a PCR, and the reaction solution held in the reaction chamber is a PCR reaction solution. With the reaction apparatus pertaining to this aspect, the progress of the PCR in the reaction chamber (such as whether or not the target nucleic acids have been amplified, or the amount of PCR amplification product) can be monitored in real time, while the PCR is conducted in less time by rapidly controlling the temperature of the PCR reaction solution.  
      The reaction apparatus pertaining to this aspect makes possible the automation of a series of operations comprising the preparation of samples containing target nucleic acids (such as extraction of nucleic acids from cells), amplification of these target nucleic acids by PCR, and monitoring (detection, measurement, qualitative analysis, quantitative analysis, etc.) of the progress of the PCR (such as whether or not the target nucleic acids have been amplified, or the amount of PCR amplification product).  
      (41) In order to achieve the stated objects, the method of the present invention comprises the steps of (a) bringing the reaction solution held in the reaction chamber into contact with a contact member, (b) controlling the temperature of the reaction solution through the contact surface between the reaction solution and the reaction chamber and/or the contact surface between the reaction solution and the contact member, (c) irradiating the reaction solution with light through the contact surface between the reaction solution and the reaction chamber and/or the contact surface between the reaction solution and the contact member, and (d) detecting fluorescent light emitted from the reaction solution through the contact surface between the reaction solution and the reaction chamber and/or the contact surface between the reaction solution and the contact member.  
      With the method of the present invention, step (b) is preferably performed after step (a). This allows the temperature of the reaction solution to be controlled rapidly through the contact surface between the reaction solution and the reaction chamber and the contact surface between the reaction solution and the contact member. The control of the reaction solution temperature through the contact surface between the reaction solution and the reaction chamber in step (b) can be carried out before step (a) or simultaneously with step (a).  
      With the method of the present invention, step (c) and step (d) are preferably performed after step (a). This allows the progress of the reaction to be monitored while the reaction proceeds by rapidly controlling the temperature of the reaction solution. The irradiation of the reaction solution with light through the contact surface between the reaction solution and the reaction chamber in step (c), and the detection of the fluorescent light emitted from the fluorescent light through the contact surface between the reaction solution and the reaction chamber in step (d) can be carried out before step (a) or simultaneously with step (a).  
      Also, with the method of the present invention, step (b), step (c), and step (d) are preferably carried out simultaneously. This allows the progress of the reaction to be monitored in real time while the reaction proceeds by rapidly controlling the reaction solution temperature.  
      The method of the present invention can be implemented, for example, by using the reaction vessel of the present invention or the reaction apparatus of the present invention.  
      (42) In a first aspect of the method of the present invention, the contact surface of the reaction chamber utilized for controlling the temperature of the reaction solution is different from the contact surface of the reaction chamber utilized for irradiating the reaction solution with light and/or the contact surface of the reaction chamber utilized for detecting fluorescent light from the reaction solution.  
      In the reaction pertaining to this aspect, of the surface where the reaction solution is in contact with the reaction solution (the contact surface of the reaction chamber), that surface utilized for controlling the temperature of the reaction solution (the surface utilized in step (b)) is separate from the surface utilized for monitoring the reaction progress (the surface utilized in step (c) and/or step (d)), which allows the temperature of the reaction solution to be controlled rapidly and also allows the region where the reaction progress is monitored to be set freely. It is also possible to monitor the reaction progress for the entire reaction solution.  
      (43) In a second aspect of the method of the present invention, a nozzle tip fitting space, into which a nozzle tip mounted on a nozzle capable of the intake and discharge of a liquid can be fitted, is formed in the contact member, and the method further comprises the steps of (e) forming a through-hole that communicates with the outside of the reaction chamber, the inside of the reaction chamber, and the nozzle tip fitting space by means of a puncture needle provided to the outside of the reaction chamber after completion of a reaction in the reaction chamber, (f) mounting the nozzle tip mounted to the nozzle in the nozzle tip fitting space, (g) bringing the outside of the reaction chamber into contact with a liquid, and (h) extracting the reaction solution held in the reaction chamber into the liquid by operating the nozzle and performing the intake and discharge of the liquid through the through-hole.  
      With the method pertaining to this aspect, steps (e), (f), and (g) can be performed in any order desired. Step (e) is performed after completion of the reaction in the reaction chamber, but steps (f) and (g) may be performed either before the reaction in the reaction chamber (including both before the reaction commences and during the course of the reaction), or may be performed after the reaction is complete. When the second reaction apparatus pertaining to the fourth aspect is utilized, of steps (e), (f), and (g), step (f) is performed first, and steps (e) and (g) are performed in any order desired. Step (h) is performed after steps (e), (f), and (g) have been performed.  
      (44) In a third aspect of the method of the present invention, the reaction occurring in the reaction chamber is a PCR.  
      With the method pertaining to this aspect, the reaction solution held in the reaction chamber is a PCR reaction solution. With the method pertaining to this aspect, the progress of the PCR in the reaction chamber (such as whether or not the target nucleic acids have been amplified, or the amount of PCR amplification product) can be monitored in real time, while the PCR is conducted in less time by rapidly controlling the temperature of the PCR reaction solution.  
    
    
     BRIEF DESCRIPTION OF THE DRAWINGS  
       FIG. 1  is a cross section illustrating a first embodiment of the reaction vessel pertaining to the present invention;  
       FIG. 2  is a top view of the reaction vessel main body of the reaction vessel pertaining to the first embodiment;  
       FIG. 3  is a bottom view of the cover member pertaining to the first embodiment;  
       FIG. 4  is a cross section illustrating a state in which the cover member is mounted on the reaction vessel main body in the reaction vessel pertaining to the first embodiment;  
       FIG. 5  is a simplified partial cross section illustrating a first embodiment of the reaction apparatus pertaining to the present invention;  
      FIGS.  6 ( i ) to ( iii ) are diagrams illustrating example layouts of the optical fibers ( FIG. 6 ( ii ) corresponds to an A-A cross section of  FIG. 5 );  
       FIG. 7  is a cross section illustrating a second embodiment of the reaction vessel pertaining to the present invention;  
       FIG. 8 ( i ) is a cross section illustrating a state in which the cover member is mounted on the reaction vessel main body in the reaction vessel pertaining to the second embodiment, and  FIG. 8 ( ii ) is a cross section illustrating a state in which the nozzle tip has been mounted on the cover member covering the reaction vessel main body in the reaction vessel pertaining to the second embodiment;  
       FIG. 9  is a partial cross section illustrating a second embodiment of the reaction apparatus pertaining to the present invention;  
       FIG. 10 ( i ) is an exploded oblique view illustrating the structure of the first temperature controller and second temperature controller provided to the reaction apparatus pertaining to the second embodiment, and  FIG. 10 ( ii ) is an oblique view illustrating the state of the first temperature controller and second temperature controller during a reaction;  
       FIG. 11  is a cross section illustrating the state near the reaction vessel during a reaction in the reaction apparatus pertaining to the second embodiment; and  
       FIG. 12  is a partial cross section illustrating the operation up to the reaction product extraction of the reaction apparatus pertaining to the second embodiment. 
    
    
     BEST MODE FOR CARRYING OUT THE INVENTION  
      Embodiments of the present invention will now be described through reference to the drawings.  
     First Embodiment  
       FIG. 1  is a cross section illustrating a first embodiment of the reaction vessel pertaining to the present invention,  FIG. 2  is a top view of the reaction vessel main body of the reaction vessel pertaining to the first embodiment,  FIG. 3  is a bottom view of the cover member pertaining to the first embodiment,  FIG. 4  is a cross section illustrating a state in which the cover member is mounted on the reaction vessel main body in the reaction vessel pertaining to the first embodiment,  FIG. 5  is a simplified partial cross section illustrating a first embodiment of the reaction apparatus pertaining to the present invention, and FIGS.  6 ( i ) to ( iii ) are diagrams illustrating example layouts of the optical fibers.  
      As shown in  FIGS. 1 and 4 , the reaction vessel  1   a  pertaining to this embodiment comprises a reaction vessel main body  2   a  and a cover member  3   a.    
      As shown in  FIGS. 1 and 2 , the reaction vessel main body  2   a  has a bottom plate  22   a  that is quadrangular in plan view, a first side plate  23   a  in the form of an angular cylinder that rises up from the edges of the bottom plate  22   a  while maintaining the same diameter, a second side plate  24   a  in the form of an angular cylinder that rises up from the top edge of the first side plate  23   a  while gradually expanding in diameter, a third side plate  25   a  in the form of an angular cylinder that rises up from the top edge of the second side plate  24   a  while maintaining the same diameter, and a lip  26   a  provided at the top edge of the third side plate  25   a.    
      As shown in  FIG. 2 , a lateral cross section of the first side plate  23   a , second side plate  24   a , and third side plate  25   a  is quadrangular, with an inner peripheral surface  213   a  of the first side plate  23   a , an inner peripheral surface  214   a  of the second side plate  24   a , and an inner peripheral surface  215   a  of the third side plate  25   a  each consisting of four planes. This quadrangular shape includes both rectangular and square.  
      As shown in  FIG. 1 , a concave component  261   a  is provided to the lip  26   a  of the reaction vessel main body  2   a , and when the cover member  3   a  is mounted on the reaction vessel main body  2   a , as shown in  FIG. 4 , the concave component  261   a  of the reaction vessel main body  2   a  fits together with a convex component  37   a  of the cover member  3   a , so that the cover member  3   a  is fixed to the reaction vessel main body  2   a.    
      As shown in  FIG. 1 , the reaction vessel main body  2   a  is equipped with a reaction chamber  21   a  that has an opening  211   a  at its upper end and is capable of holding the reaction solution  4   a . As shown in  FIG. 1 , the reaction chamber  21   a  is formed in the reaction vessel main body  2   a  as a concave component having an opening  211   a  at its upper end. The reaction chamber  21   a  is a concave component formed by the bottom plate  22   a , the first side plate  23   a , the second side plate  24   a , and the third side plate  25   a . The upper surface  212   a  of the bottom plate  22   a  corresponds to the bottom surface of the reaction chamber  21   a , and the inner peripheral surface  231   a  of the first side plate  23   a , the inner peripheral surface  214   a  of the second side plate  24   a , and the inner peripheral surface  215   a  of the third side plate  25   a  correspond to the inner peripheral surface of the reaction chamber  21   a.    
      As shown in  FIG. 1 , the opening area of the opening  211   a  of the reaction chamber  21   a  is somewhat larger than the surface area of the bottom surface of the reaction chamber  21   a , and the structure is such that the reaction solution  4   a  added through the opening  211   a  easily reaches the bottom surface of the reaction chamber  21   a  all by itself (without any downward force other than gravity being applied to the reaction solution  4   a ). Depending on how the reaction solution  4   a  is added, it may adhere to the inner peripheral surface of the reaction chamber  21   a , in which case the reaction solution  4   a  can be made to reach the bottom of the reaction chamber  21   a  by using a vortex mixer or the like to vibrate the reaction vessel main body  2   a.    
      As shown in  FIG. 1 , the bottom plate  22   a , the first side plate  23   a , the second side plate  24   a , and the third side plate  25   a  have a substantially uniform thickness. “Substantially uniform thickness” includes a thickness that is uniform. The thickness of the various plates can be varied as needed, but the bottom plate  22   a  is preferably a thin plate from the standpoint of rapidly controlling the temperature of the reaction solution  4   a  held in the reaction chamber  21   a . Also, it is preferable for the first side plate  23   a  to be a thin plate because it will be easier to set the conditions for irradiating the reaction solution  4   a  held in the reaction chamber  21   a  with excitation light and the conditions for detecting the fluorescent light emitted from the reaction solution  4   a . The thickness of the thin plates can be suitably determined according to the material of which the thin plates are made and other factors, but about 0.1 to 0.5 mm is preferable in the case of a plastic, for example.  
      As shown in  FIG. 1 , the reaction vessel main body  2   a  is only equipped with one reaction chamber  21   a , but the number of reaction chambers and their location in the reaction vessel main body can be varied as needed. For instance, the reaction vessel main body may be equipped with eight reaction chambers aligned in a row, or may be equipped with 8 longitudinal rows×12 lateral rows for a total of 96 reaction chambers. Sample processing can be carried out more efficiently if the reaction vessel main body is equipped with a plurality of reaction chambers. For example, a sample dispensing apparatus comprising an eight-head nozzle unit is commercially available, if the reaction vessel main body is equipped with eight reaction chambers in a row, this sample dispensing apparatus can be utilized to automate the dispensing of the reaction solution into the reaction chambers.  
      As shown in  FIG. 1 , when the reaction vessel main body  2   a  is not yet covered by the cover member  3   a , the reaction solution  4   a  held in the reaction chamber  21   a  is in contact with the bottom surface and the inner peripheral surface of the reaction chamber  21   a.    
      As shown in  FIG. 1 , the cover member  3   a  has a convex component  31   a  protruding downward, and a flat plate  36   a  provided to the top end of the convex component  31   a.    
      As shown in  FIG. 1 , a lip protruding downward is provided to the peripheral edge of the flat plate  36   a , and a convex component  37   a  that protrudes in the direction of the convex component  31   a  is provided to the bottom end of this lip. When the reaction vessel main body  2   a  is covered by the cover member  3   a , as shown in  FIG. 4 , the convex component  37   a  of the cover member  3   a  fits into the concave component  261   a  of the reaction vessel main body  2   a  so that the cover member  3   a  is fixed to the reaction vessel main body  2   a.    
      As shown in  FIGS. 1 and 3 , the convex component  31   a  comprises the pressing part  32   a  comprising a flat plate that is quadrangular in plan view, a first side plate  33   a  in the form of an angular cylinder that rises up from the edges of the pressing part  32   a  so as to gradually increase in diameter, a second side plate  34   a  in the form of an angular cylinder that rises up from the top end of the first side plate  33   a  so as to gradually increase in diameter, and a third side plate  35   a  in the form of an angular cylinder that rises up from the top end of the second side plate  34   a  so as to maintain the same diameter. The convex component  31   a  is contiguous with the flat plate  36   a  at the top end of the third side plate  35   a.    
      As shown in  FIG. 3 , a lateral cross section of the first side plate  33   a , the second side plate  34   a , and the third side plate  35   a  is quadrangular, with an outer peripheral surface  313   a  of the first side plate  33   a , an outer peripheral surface  314   a  of the second side plate  34   a , and an outer peripheral surface  315   a  of the third side plate  35   a  each consisting of four planes. This quadrangular shape includes both rectangular and square.  
      As shown in  FIG. 1 , the pressing part  32   a , the first side plate  33   a , the second side plate  34   a , and the third side plate  35   a  have a substantially uniform thickness. “Substantially uniform thickness” includes a thickness that is uniform. The thickness of the various plates can be varied as needed, but the pressing part  32   a  is preferably a thin plate from the standpoint of rapidly controlling the temperature of the reaction solution  4   a  held in the reaction chamber  21   a . The thickness of the thin plates can be suitably determined according to the material of which the thin plates are made and other factors, but about 0.1 to 0.5 mm is preferable in the case of a plastic, for example.  
      As shown in  FIGS. 1 and 3 , the cover member  3   a  only has one convex component  31   a , but the number of convex components and their location in the cover member can be varied as needed according to the number and locations of the reaction chambers had by the reaction vessel main body.  
      The convex component  31   a  is provided to the cover member  3   a  so as to fit into the reaction chamber  21   a  formed as a concave component in the reaction vessel main body  2   a , and when the reaction vessel main body  2   a  is covered by the cover member  3   a , as shown in  FIG. 4 , the opening  211   a  of the reaction chamber  21   a  is sealed off by the cover member  3   a.    
      The convex component  31   a  is provided so that the lower surface  312   a  of the pressing part  32   a  of the convex component  31   a  does not come into contact with the bottom surface of the reaction chamber  21   a  when the cover member  3   a  is mounted on the reaction vessel main body  2   a . Therefore, when the cover member  3   a  covers the reaction vessel main body  2   a , as shown in  FIG. 4 , a gap (tightly closed space S 1   a ) is formed between the lower surface  312   a  of the pressing part  32   a  of the convex component  31   a  and the bottom surface of the reaction chamber  21   a.    
      The pressing part  32   a  of the convex component  31   a  is provided so as to be opposed against the bottom plate  22   a  of the reaction vessel main body  2   a  when the cover member  3   a  is mounted on the reaction vessel main body  2   a , and when the cover member  3   a  is placed over the reaction vessel main body  2   a , as shown in  FIG. 4 , the reaction solution  4   a  is held in the form of a thin layer between the lower surface  312   a  (contact surface of the cover member) of the pressing part  32   a  of the convex component  31   a  and the upper surface  212   a  (opposing surface of the reaction chamber) of the bottom plate  22   a  of the reaction vessel main body  2   a.    
      At this point, the reaction solution  4   a  present in the form of a thin layer between the lower surface  312   a  (contact surface of the cover member) of the pressing part  32   a  of the convex component  31   a  and the upper surface  212   a  (opposing surface of the reaction chamber) of the bottom plate  22   a  of the reaction vessel main body  2   a  is in a state of being enveloped by the inner peripheral surface  213   a  (enveloping surface of the reaction chamber) of the first side plate  23   a  of the reaction vessel main body  2   a . Specifically, when the cover member  3   a  is placed over the reaction vessel main body  2   a , as shown in  FIG. 4 , the tightly closed space S 1   a  is formed by the lower surface  312   a  (contact surface of the cover member) of the pressing part  32   a  of the convex component  31   a , the upper surface  212   a  (opposing surface of the reaction chamber) of the bottom plate  22   a  of the reaction vessel main body  2   a , and the inner peripheral surface  213   a  (enveloping surface of the reaction chamber) of the first side plate  23   a  of the reaction vessel main body  2   a , and part of the reaction solution  4   a  is held in the form of a thin layer in this tightly closed space S 1   a.    
      As shown in  FIG. 4 , the reaction solution  4   a  held in the tightly closed space S 1   a  is in contact with the lower surface  312   a  (contact surface of the cover member) of the pressing part  32   a  of the convex component  31   a , the upper surface  212   a  (opposing surface of the reaction chamber) of the bottom plate  22   a  of the reaction vessel main body  2   a , and the inner peripheral surface  213   a  (enveloping surface of the reaction chamber) of the first side plate  23   a  of the reaction vessel main body  2   a.    
      The pressing part  32   a  of the convex component  31   a  is provided so as to press on the reaction solution  4   a  held in the reaction chamber  21   a  in the course of the cover member  3   a  being placed over the reaction vessel main body  2   a , and this pressing by the pressing part  32   a  gradually pushes the reaction solution  4   a  to the upper part of the reaction chamber  21   a . When the cover member  3   a  then covers the reaction vessel main body  2   a , as shown in  FIG. 4 , the outer peripheral surface  313   a  of the first side plate  33   a  of the cover member  3   a  strikes the inner peripheral surface  214   a  of the second side plate  24   a  of the reaction vessel main body  2   a . This limits the downward movement of the pressing part  32   a , so that the distance remains constant between the lower surface  312   a  (contact surface of the cover member) of the pressing part  32   a  of the convex component  31   a  and the upper surface  212   a  (opposing surface of the reaction chamber) of the bottom plate  22   a  of the reaction vessel main body  2   a . In this embodiment, the surface (abutting surface) when the cover member  3   a  and the reaction vessel main body  2   a  come together is in tapered form, but the abutting surface can also be, for example, a surface perpendicular to the direction in which the cover member is placed over the reaction vessel main body.  
      When the cover member  3   a  covers the reaction vessel main body  2   a , as shown in  FIG. 4 , the outer peripheral surface  313   a  of the first side plate  33   a  of the cover member  3   a  fits snugly against the inner peripheral surface  214   a  of the second side plate  24   a  of the reaction vessel main body  2   a , and the outer peripheral surface  315   a  of the third side plate  35   a  of the cover member  3   a  also fits snugly against the inner peripheral surface  215   a  of the third side plate  25   a  of the reaction vessel main body  2   a . This keeps the tightly closed space S 1   a  tightly closed, and also prevents contamination of the reaction solution  4   a  held in the tightly closed space S 1   a.    
      On the other hand, even when the reaction vessel main body  2   a  is covered by the cover member  3   a , as shown in FIG.  4 , the second side plate  34   a  of the cover member  3   a  does not fit snugly against the second side plate  24   a  or third side plate  25   a  of the reaction vessel main body  2   a . Specifically, a tightly closed space S 2   a  (surplus reaction solution holder) for holding any surplus reaction solution  4   a  that will not be held in the tightly closed space S 1   a  is formed between the second side plate  34   a  of the cover member  3   a  and the second side plate  24   a  and third side plate  25   a  of the reaction vessel main body  2   a.    
      When the pressing part  32   a  presses against the reaction solution  4   a , any air inside the reaction chamber  21   a , bubbles in the reaction solution  4   a , and so forth are pushed along with the reaction solution  4   a  to the top part of the reaction chamber  21   a  and held in the tightly closed space S 2   a , and part thereof is discharged to outside the reaction chamber  21   a , which prevents the admixture of air into the tightly closed space S 1   a  and the admixture of bubbles into the reaction solution  4   a  held in the tightly closed space S 1   a.    
      The convex component  31   a  is provided so that the reaction solution  4   a  present between the lower surface  312   a  (contact surface of the cover member) of the pressing part  32   a  of the convex component  31   a  and the upper surface  212   a  (opposing surface of the reaction chamber) of the bottom plate  22   a  of the reaction vessel main body  2   a  is in the form of a thin layer when the cover member  3   a  is mounted on the reaction vessel main body  2   a . Specifically, the convex component  31   a  is provided so that the distance between the lower surface  312   a  (contact surface of the cover member) of the pressing part  32   a  of the convex component  31   a  and the upper surface  212   a  (opposing surface of the reaction chamber) of the bottom plate  22   a  of the reaction vessel main body  2   a  is shortened when the cover member  3   a  is placed over the reaction vessel main body  2   a.    
      The distance between the lower surface  312   a  (contact surface of the cover member) of the pressing part  32   a  of the convex component  31   a  and the upper surface  212   a  (opposing surface of the reaction chamber) of the bottom plate  22   a  of the reaction vessel main body  2   a  (that is, the thickness of the thin layer of reaction solution  4   a ) is preferably 0.1 to 0.5 mm. Also, distance between the lower surface  312   a  (contact surface of the cover member) of the pressing part  32   a  of the convex component  31   a  and the upper surface  212   a  (opposing surface of the reaction chamber) of the bottom plate  22   a  of the reaction vessel main body  2   a  (that is, the thickness of the thin layer of reaction solution  4   a ) is preferably the same at all locations.  
      The reaction vessel main body  2   a  and the cover member  3   a  are made of a material that will not be corroded by the reaction solution  4   a , that can withstand the conditions of the reaction that occurs in the reaction chamber  21   a  (such as the reaction temperature), and that has optical transparency.  
      Since the reaction vessel main body  2   a  is made entirely of a light transmitting material, light can be transmitted from the outside of the reaction vessel main body  2   a  to the reaction solution  4   a  held in the tightly closed space S 1   a , and from the reaction solution  4   a  held in the tightly closed space S 1   a  to the outside of the reaction vessel main body  2   a , through the bottom plate  22   a  and the first side plate  23   a  of the reaction vessel main body  2   a.    
      Also, since the cover member  3   a  is made entirely of a light transmitting material, light can be transmitted from the outside of the reaction vessel main body  2   a  to the reaction solution  4   a  held in the tightly closed space S 1   a , and from the reaction solution  4   a  held in the tightly closed space S 1   a  to the outside of the reaction vessel main body  2   a , through the pressing part  32   a  of the cover member  3   a.    
      However, the reaction vessel main body  2   a  and the cover member  3   a  do not need to be made entirely from a light transmitting material, and it will be sufficient if the portion where light needs to be transmitted in order to monitor the progress of the reaction occurring in the reaction chamber  21   a  is made from a light transmitting material.  
      For example, if the irradiation of the reaction solution  4   a  with the excitation light and the detection of fluorescent light from the reaction solution  4   a  are performed through the first side plate  23   a  of the reaction vessel main body  2   a , then just the first side plate  23   a  of the reaction vessel main body  2   a  may be made of a light transmitting material. If the irradiation of the reaction solution  4   a  with the excitation light and the detection of fluorescent light from the reaction solution  4   a  are performed through the bottom plate  22   a  of the reaction vessel main body  2   a , then just the bottom plate  22   a  of the reaction vessel main body  2   a  may be made of a light transmitting material. If the irradiation of the reaction solution  4   a  with the excitation light and the detection of fluorescent light from the reaction solution  4   a  are performed through the pressing part  32   a  of the cover member  3   a , then just the pressing part  32   a  of the cover member  3   a  may be made of a light transmitting material.  
      Also, just the reaction vessel main body  2   a  or the cover member  3   a  may be made of a light transmitting material, and the other made of an opaque material. For example, if the irradiation of the reaction solution  4   a  with the excitation light and the detection of fluorescent light from the reaction solution  4   a  are performed through the first side plate  23   a  of the reaction vessel main body  2   a , the cover member  3   a  may be made of a light transmitting material.  
      Examples of the material of the reaction vessel main body  2   a  and the cover member  3   a  include transparent or semitransparent thermoplastic resins and glass. If a thermoplastic resin is selected as the material of the reaction vessel main body  2   a  and the cover member  3   a , the reaction vessel main body  2   a  and the cover member  3   a  can be easily formed by a standard process such as injection molding. If the reaction will reach a high temperature (such as 90 to 100° C.), then it is preferable to use a material with excellent heat resistance, such as an engineering plastic.  
      As shown in  FIG. 5 , the reaction apparatus  10   a  pertaining to this embodiment comprises the reaction vessel  1   a  supported on a seat  53   a , a temperature controller  6   a  equipped with thermoelectric semiconductor elements  61   a  and  62   a , a light source  7   a , a fluorescent light detector  8   a , and a plurality of optical fibers  9   a.    
      As shown in  FIG. 5 , the thermoelectric semiconductor element  61   a  of the temperature controller  6   a  is attached via a heat-conducting metal plate  51   a  to the pressing part  32   a  (upper surface of the pressing part  32   a ) of the cover member  3   a , and the thermoelectric semiconductor element  62   a  is attached via the heat-conducting metal plate  51   a  to the bottom plate  22   a  (lower surface of the bottom plate  22   a ) of the reaction vessel main body  2   a . The thermoelectric semiconductor elements are types that can be utilized as cooling elements and/or as heating elements, an example of which is a Peltier element.  
      The temperature controller  6   a  is designed so as to allow control of the heating and cooling performed by the thermoelectric semiconductor elements  61   a  and  62   a , and the thermoelectric semiconductor elements  61   a  and  62   a  are electrically connected to the temperature controller  6   a . Also, as shown in  FIG. 5 , heat radiators  52   a  having cooling fins are mounted on the thermoelectric semiconductor elements  61   a  and  62   a , allowing the forced cooling of the thermoelectric semiconductor elements  61   a  and  62   a . With the temperature controller  6   a , the temperature of the reaction solution  4   a  held in the tightly closed space S 1   a  of the reaction vessel  1   a  can be rapidly controlled by the movement of heat through the pressing part  32   a  of the cover member  3   a  and by the movement of heat through the bottom plate  22   a  of the reaction vessel main body  2   a.    
      The reaction solution  4   a  is a PCR-use reaction solution, and the amount of the reaction solution  4   a  held in the tightly closed space S 1   a  is preferable about 2 to 50 μL. A PCR proceeds as the temperature of the reaction solution  4   a  is controlled by the temperature controller  6   a . Here, since the reaction solution  4   a  is held in the tightly closed space S 1   a  in the form of a thin layer, the ratio of surface area to volume is higher, and furthermore nearly all of this surface area is accounted for by the upper and lower surfaces of the thin layer, that is, by the lower surface  312   a  (contact surface of the cover member) of the pressing part  32   a  of the cover member  3   a  and the upper surface  212   a  (opposing surface of the reaction chamber) of the bottom plate  22   a  of the reaction vessel main body  2   a . Therefore, the temperature of the reaction solution  4   a  held in the tightly closed space S 1   a  can be rapidly controlled by the movement of heat through the pressing part  32   a  of the cover member  3   a  and by the movement of heat through the bottom plate  22   a  of the reaction vessel main body  2   a , which means that the PCR takes less time.  
      The light source  7   a  is an apparatus capable of emitting excitation light that will excite the fluorescent dye contained in the reaction solution  4   a . As shown in  FIG. 5 , the plurality of optical fibers  9   a  are mounted in the light source  7   a , and excitation light emitted from the light source  7   a  is directed through the optical fibers  9   a . The optical fibers  9   a  are disposed around the first side plate  23   a  (the outer peripheral surface of the first side plate  23   a ) of the reaction vessel main body  2   a  as shown  FIG. 5  and  FIG. 6 ( ii ), and the excitation light emitted from the light source  7   a  and through the optical fibers  9   a  irradiates the reaction solution  4   a  held in the tightly closed space S 1   a  through the first side plate  23   a  of the reaction vessel main body  2   a.    
      In addition to H 2 O, buffers, MgCl 2 , dNTP mixes, primers, template DNA, Taq polymerase, and so forth, the reaction solution  4   a  also contains ethidium bromide, SYBR Green I, Pico Green, and other such fluorescent dyes that can serve as an index of the progress of a PCR (such as whether or not the target nucleic acids have been amplified, or the amount of PCR amplification product). Therefore, when the reaction solution  4   a  held in the tightly closed space S 1   a  is irradiated with the excitation light, these fluorescent dyes emit fluorescent light. The fluorescent light emitted from the reaction solution  4   a  held in the tightly closed space S 1   a  is transmitted through the first side plate  23   a  of the reaction vessel main body  2   a  to the outside of the reaction vessel main body  2   a.    
      The fluorescent light detector  8   a  is an apparatus capable of detecting fluorescent light emitted from the reaction solution  4   a . As shown in  FIG. 5 , the plurality of optical fibers  9   a  are mounted to the fluorescent light detector  8   a , and the fluorescent light that is emitted from the reaction solution  4   a  held in the tightly closed space S 1   a  and transmitted through the first side plate  23   a  of the reaction vessel main body  2   a  to outside of the reaction vessel main body  2   a  is received through the optical fibers  9   a  and detected by the fluorescent light detector  8   a.    
      There are no particular restrictions on the structure of the light source  7   a  and the fluorescent light detector  8   a , and any standard apparatus equipped with filters, reflecting mirrors, lenses, and so forth can be used.  
      The fluorescent intensity of the light emitted from the reaction solution  4   a  is proportional to the amount of DNA contained in the reaction solution  4   a , so the progress of the PCR (such as whether or not the target nucleic acids have been amplified, or the amount of PCR amplification product) can be monitored in real time (that is, instantly during the course of the PCR) by detecting the fluorescent intensity.  
      The optical fibers  9   a  are connected at one end to the light source  7   a  or the fluorescent light detector  8   a , and at the other end are disposed around the first side plate  23   a  (the outer peripheral surface of the first side plate  23   a ) of the reaction vessel main body  2   a.    
       FIG. 6  shows examples of the layout of the optical fibers  9   a .  FIG. 6  corresponds to an A-A cross section of  FIG. 5 .  
      In  FIG. 6 ( i ), optical fibers  9   a  are disposed at one end on one side of the outer peripheral surface of the first side plate  23   a , which is quadrangular in cross sectional shape, so that irradiation of the reaction solution  4   a  with excitation light and the reception of the fluorescent light emitted from the reaction solution  4   a  can both be performed by the optical fibers  9   a.    
      In  FIG. 6  ( ii ), optical fibers  9   a  are disposed at one end on two facing sides of the outer peripheral surface of the first side plate  23   a , which is quadrangular in cross sectional shape, so that irradiation of the reaction solution  4   a  with excitation light is performed by the optical fibers  9   a  disposed on one side, and the reception of the fluorescent light emitted from the reaction solution  4   a  is performed by the optical fibers  9   a  disposed on the other side.  
      In  FIG. 6 ( iii ), optical fibers  9   a  are disposed at one end on two perpendicular sides of the outer peripheral surface of the first side plate  23   a , which is quadrangular in cross sectional shape, so that irradiation of the reaction solution  4   a  with excitation light is performed by the optical fibers  9   a  disposed on one side, and the reception of the fluorescent light emitted from the reaction solution  4   a  is performed by the optical fibers  9   a  disposed on the other side.  
      With the examples shown in FIGS.  6 ( i ) to ( iii ), the optical fibers are disposed perpendicular to the outer peripheral surface of the first side plate  23   a , which simplifies the setting of the irradiation conditions and light reception conditions.  
      With the reaction apparatus  10   a , of the reaction vessel main body  2   a  and the cover member  3   a , the portions utilized for controlling the temperature of the reaction solution (the pressing part  32   a  of the cover member  3   a  and the bottom plate  22   a  of the reaction vessel main body  2   a ) are separate from the portions utilized for monitoring the progress of the reaction (the first side plate  23   a  of the reaction vessel main body  2   a ), which allows the temperature of the reaction solution  4   a  to be controlled rapidly and also allows the region where the reaction progress is monitored to be set freely.  
      With the reaction apparatus  10   a , the type of irradiating excitation light and the type of detected fluorescent light may be the same among the various optical fibers, or may vary with each optical fiber or by optical fiber group. If the reaction solution  4   a  is made ahead of time to contain a plurality of different fluorescent dyes, and irradiation with excitation light corresponding to each fluorescent dye and the detection of the fluorescent light emitted from each fluorescent dye are performed by individual optical fiber or by optical fiber group, then different PCRs can be conducted at the same time, and the progress of the reactions (such as whether or not the target nucleic acids have been amplified, or the amount of PCR amplification product) can be monitored in real time. Also, if the same excitation light is emitted and the same fluorescent light detected by a plurality of optical fibers, then the entire reaction solution  4   a  can be irradiated with excitation light and the progress of the reaction can be monitored for the entire reaction solution  4   a  by disposing the optical fibers over one entire side of the outer peripheral surface of the first side plate  23   a , which is quadrangular in lateral cross sectional shape.  
      The first embodiment described above is given in order to facilitate an understanding of the present invention, and does not limit the present invention in any way. Therefore, the various elements disclosed in the first embodiment should be construed as encompassing all design modifications, equivalents, etc., within the technological scope of the present invention.  
      For example, the optical fibers  9   a  can be disposed on the upper surface of the pressing part  32   a  of the cover member  3   a . In this case, the irradiation of the reaction solution  4   a  with the excitation light and/or the detection of the fluorescent light from the reaction solution  4   a  can be performed through the pressing part  32   a  of the cover member  3   a.    
      The optical fibers  9   a  can also be disposed on the lower surface of the bottom plate  22   a  of the reaction vessel main body  2   a . In this case, the irradiation of the reaction solution  4   a  with the excitation light and/or the detection of the fluorescent light from the reaction solution  4   a  can be performed through the bottom plate  22   a  of the reaction vessel main body  2   a.    
      Also, the irradiation of the reaction solution  4   a  with the excitation light and the detection of the fluorescent light from the reaction solution  4   a  can be performed by utilizing lenses or the like, rather than optical fibers. The use of lenses facilitates irradiating the entire reaction solution  4   a  with excitation light and the detection of fluorescent light emitted from the entire reaction solution  4   a.    
     Second Embodiment  
       FIG. 7  is a cross section illustrating a second embodiment of the reaction vessel pertaining to the present invention;  FIG. 8 ( i ) is a cross section illustrating a state in which the cover member is mounted on the reaction vessel main body in the reaction vessel pertaining to the second embodiment;  FIG. 8 ( ii ) is a cross section illustrating a state in which the nozzle tip has been mounted on the cover member covering the reaction vessel main body in the reaction vessel pertaining to the second embodiment;  FIG. 9  is a partial cross section illustrating a second embodiment of the reaction apparatus pertaining to the present invention;  FIG. 10 ( i ) is an exploded oblique view illustrating the structure of the first temperature controller and second temperature controller provided to the reaction apparatus pertaining to the second embodiment;  FIG. 10 ( ii ) is an oblique view illustrating the state of the first temperature controller and second temperature controller during a reaction;  FIG. 11  is a cross section illustrating the state near the reaction vessel during a reaction in the reaction apparatus pertaining to the second embodiment; and  FIG. 12  is a partial cross section illustrating the operation up to the reaction product extraction of the reaction apparatus pertaining to the second embodiment.  
      As shown in  FIGS. 7 and 8 , the reaction vessel  1   b  pertaining to this embodiment comprises a reaction vessel main body  2   b  and a cover member  3   b.    
      As shown in  FIG. 7 , the reaction vessel main body  2   b  has a disk-shaped bottom plate  22   b , a first side plate  23   b  in the form of a cylinder that rises up from the edges of the bottom plate  22   b  while maintaining the same diameter, a tapering second side plate  24   b  that rises up from the top edge of the first side plate  23   b  while gradually expanding in diameter, a third side plate  25   b  in the form of a cylinder that rises up from the top edge of the second side plate  24   b  while maintaining the same diameter, and a flange  26   b  provided at the top edge of the third side plate  25   b.    
      The bottom plate  22   b  and the first to third side plates  23   b  to  25   b  of the reaction vessel main body  2   b  consist of thin plates made of a material that will not be corroded by the reaction solution, that can withstand the conditions of the reaction that occurs in the reaction chamber (such as the reaction temperature), and that has optical transparency. The thickness of the thin plates is preferably about 0.1 to 0.5 mm.  
      As shown in  FIG. 7 , a reaction chamber  20   b  surrounded by the bottom plate  22   b  and the first to third side plates  23   b  to  25   b  is formed in the reaction vessel main body  2   b , and an opening  21   b  that leads to the reaction chamber  20   b  is formed at the top end of the reaction vessel main body  2   b.    
      The reaction chamber  20   b  is designed so that a reaction solution can be introduced through the opening  21   b . Also, the reaction chamber  20   b  does not lead to any opening other than the opening  21   b , and is therefore closed off when the opening  21   b  is sealed (see  FIG. 8 ( i )).  
      The inside diameter of the third side plate  25   b  of the reaction vessel main body  2   b  is substantially the same as the outside diameter of a second side plate  34   b  of the cover member  3   b , so that when the cover member  3   b  is placed over the reaction vessel main body  2   b , the inner peripheral surface of the third side plate  25   b  of the reaction vessel main body  2   b  fits snugly against the outer peripheral surface of the second side plate  34   b  of the cover member  3   b  (see  FIG. 8 ( i )).  
      As shown in  FIG. 7 , a convex component  27   b  is provided on the inner peripheral surface of the third side plate  25   b  of the reaction vessel main body  2   b , and the convex component  27   b  is designed to fit into a concave component  36   b  provided on the outer peripheral surface of the second side plate  34   b  of the cover member  3   b  (see  FIG. 8 ( i )).  
      As shown in  FIG. 7 , an abutting surface  28   b  is provided to the top end of the first side plate  23   b  of the reaction vessel main body  2   b , and the abutting surface  28   b  is designed so as to strike the a bottom plate  32   b  of the cover member  3   b  when the cover member  3   b  is placed over the reaction vessel main body  2   b  (see  FIG. 8 ( i )).  
      As shown in  FIG. 7 , the cover member  3   b  has the disk-shaped bottom plate  32   b , a tapering first side plate  33   b  that rises up from the edges of the bottom plate  32   b  so as to gradually increase in diameter, the second side plate  34   b  in the form of a cylinder that rises up from the top end of the first side plate  33   b  while maintaining the same diameter, and a flange  35   b  provided to the top edges of the second side plate  34   b.    
      The bottom plate  32   b , the first side plate  33   b , and the second side plate  34   b  of the cover member  3   b  consist of thin plates made of a material that will not be corroded by the reaction solution, that can withstand the conditions of the reaction that occurs in the reaction chamber (such as the reaction temperature), and that has optical transparency (such as transparent or semitransparent thermoplastic resins and glass). The thickness of the thin plates is preferably about 0.1 to 0.5 mm.  
      As shown in  FIG. 7 , a nozzle tip fitting space  30   b  surrounded by the bottom plate  32   b , the first side plate  33   b , and the second side plate  34   b  is formed in the cover member  3   b , and a nozzle tip fitting hole  31   b  that leads into the nozzle tip fitting space  30   b  is formed at the top end of the cover member  3   b.    
      The nozzle tip fitting space  30   b  is formed so that a nozzle tip  4   b  can be mounted through the nozzle tip fitting hole  31   b  (see  FIG. 8 ( ii )). Also, the nozzle tip fitting space  30   b  does not lead to any opening other than the nozzle tip fitting hole  31   b , and is therefore closed off when the nozzle tip fitting hole  31   b  is sealed (see  FIG. 8 ( ii )).  
      The nozzle tip fitting hole  31   b  is formed in the portion of the cover member  3   b  other than the portion where the opening  21   b  of the reaction vessel main body  2   b  is sealed off, so when the cover member  3   b  is mounted on the reaction vessel main body  2   b , the nozzle tip  4   b  can be mounted in the nozzle tip fitting space  30   b  through the nozzle tip fitting hole  31   b  (see  FIG. 8 ( ii )).  
      The deepest portion of the nozzle tip fitting space  30   b  (the portion of the nozzle tip fitting space  30   b  farthest away from the nozzle tip fitting hole  31   b ) is formed by the bottom plate  32   b  of the cover member  3   b , and the nozzle tip  4   b  is mounted toward the deepest part of the nozzle tip fitting space  30   b  from the nozzle tip fitting hole  31   b  (see  FIG. 8 ( ii )).  
      The mounting direction of the nozzle tip  4   b  with respect to the nozzle tip fitting space  30   b  is perpendicular or substantially perpendicular to the surface (the lower surface of the bottom plate  22   b  of the reaction vessel  1   b ) on which the reaction vessel  1   b  is placed (see  FIG. 8 ( ii )), so the force exerted on the reaction vessel  1   b  by the nozzle tip  4   b  in the mounting of the nozzle tip  4   b  is a force that is perpendicular or substantially perpendicular to the surface on which the reaction vessel  1   b  is placed. Therefore, the nozzle tip  4   b  can be easily mounted in the nozzle tip fitting space  30   b  without the reaction vessel  1   b  shifting its position while the nozzle tip  4   b  is being mounted.  
      The outside diameter of the second side plate  34   b  of the cover member  3   b  is substantially the same as the inside diameter of the third side plate  25   b  of the reaction vessel main body  2   b , so that the inner peripheral surface of the third side plate  25   b  of the reaction vessel main body  2   b  fits snugly against the outer peripheral surface of the second side plate  34   b  of the cover member  3   b  when the cover member  3   b  is placed over the reaction vessel main body  2   b  (see  FIG. 8 ( i )).  
      As shown in  FIG. 7 , the concave component  36   b  is provided on the outer peripheral surface of the second side plate  34   b  of the cover member  3   b , and the concave component  36   b  mates with the convex component  27   b  provided on the inner peripheral surface of the third side plate  25   b  of the reaction vessel main body  2   b  (see  FIG. 8 ( i )).  
      As shown in  FIG. 7 , a convex component  37   b  is provided on the inner peripheral surface of the second side plate  34   b  of the cover member  3   b , and the convex component  37   b  mates with a concave component  49   b  provided on the outer peripheral surface of a second side plate  44   b  of the nozzle tip  4   b  (see  FIG. 8 ( ii )).  
      As shown in  FIG. 7 , an abutting surface  38   b  is provided to the top end of the second side plate  34   b  of the cover member  3   b , and the abutting surface  38   b  strikes a third side plate  46   b  of the nozzle tip  4   b  when the nozzle tip  4   b  is mounted in the nozzle tip fitting space  30   b  (see  FIG. 8 ( ii )).  
      When the cover member  3   b  is placed over the reaction vessel main body  2   b , as shown in  FIG. 8 ( i ), the inner peripheral surface of the third side plate  25   b  of the reaction vessel main body  2   b  fits snugly against the outer peripheral surface of the second side plate  34   b  of the cover member  3   b , the opening  21   b  of the reaction vessel main body  2   b  is sealed off by the bottom plate  32   b , the first side plate  33   b , and the second side plate  34   b  of the cover member  3   b , and the reaction chamber  20   b  of the reaction vessel main body  2   b  is closed off. Here, as shown in  FIG. 8 ( i ), the convex component  27   b  provided to the third side plate  25   b  of the reaction vessel main body  2   b  fits into the concave component  36   b  provided to the second side plate  34   b  of the cover member  3   b , so that the cover member  3   b  is fixed to the reaction vessel main body  2   b , which makes the covering of the reaction vessel main body  2   b  by the cover member  3   b  more secure.  
      Also, when the cover member  3   b  is placed over the reaction vessel main body  2   b , as shown in  FIG. 8 ( i ), the abutting surface  28   b  provided to the top end of the first side plate  23   b  of the reaction vessel main body  2   b  strikes the bottom plate  32   b  of the cover member  3   b , which defines the location of the bottom plate  32   b  of the cover member  3   b  within the reaction chamber  20   b  (in this embodiment, the bottom plate  32   b  of the cover member  3   b  is limited so as not to come into contact with the bottom plate  22   b  of the reaction vessel main body  2   b ), and forms a tightly closed space S 1   b  between the bottom plate  22   b  of the reaction vessel main body  2   b  and the bottom plate  32   b  of the cover member  3   b . Specifically, a tightly closed space S 1   b  is formed by the lower surface (contact surface of the cover member) of the bottom plate  32   b  of the cover member  3   b , the upper surface (opposing surface of the reaction chamber) of the bottom plate  22   b  of the reaction vessel main body  2   b , and the inner peripheral surface (enveloping surface of the reaction chamber) of the first side plate  23   b  of the reaction vessel main body  2   b , so that part of the reaction solution is held in the form of a thin layer in this tightly closed space S 1   b . The reaction solution held in the tightly closed space S 1   b  is in contact with the lower surface (contact surface of the cover member) of the bottom plate  32   b  of the cover member  3   b , the upper surface (opposing surface of the reaction chamber) of the bottom plate  22   b  of the reaction vessel main body  2   b , and the inner peripheral surface (enveloping surface of the reaction chamber) of the first side plate  23   b  of the reaction vessel main body  2   b.    
      When the cover member  3   b  is placed over the reaction vessel main body  2   b , as shown in  FIG. 8 ( i ), a tightly closed space S 2   b  is formed between the second side plate  24   b  and the third side plate  25   b  of the reaction vessel main body  2   b  and the first side plate  33   b  of the cover member  3   b . The reaction solution is held in the tightly closed space S 1   b  formed within the reaction chamber  20   b  when the cover member  3   b  is put in place, and any surplus reaction solution that will not be held in the tightly closed space S 1   b  is held in the tightly closed space S 2   b . At this point the reaction solution is pressed by the bottom plate  32   b  of the cover member  3   b , any air inside the reaction chamber  20   b , bubbles in the reaction solution, and so forth are pushed along with the reaction solution to the top part of the reaction chamber  20   b  and held in the tightly closed space S 2   b , and part thereof is discharged from the opening  21   b  to outside the reaction chamber  20   b , which prevents the admixture of air into the tightly closed space S 1   b  and the admixture of bubbles into the reaction solution held in the tightly closed space S 1   b.    
      As shown in  FIG. 7 , the nozzle tip  4   b  pertaining to this embodiment has a disk-shaped distal end plate  43   b  constituting the distal end of the nozzle tip  4   b , the tapering first side plate  44   b  that rises up from the edges of the distal end plate  43   b  while gradually expanding in diameter, a cylindrical second side plate  45   b  that rises up from the top end of the first side plate  44   b  while maintaining the same diameter, a tapering third side plate  46   b  that rises up from the edges of the second side plate  45   b  while gradually expanding in diameter, a cylindrical fourth side plate  47   b  that rises up from the top end of the third side plate  46   b  while maintaining the same diameter, and a flange  48   b  provided to the top edges of the fourth side plate  47   b.    
      As shown in  FIG. 7 , an internal space  40   b  surrounded by the distal end plate  43   b  and the first to fourth side plates  44   b  to  47   b  is formed in the nozzle tip  4   b . A nozzle mounting hole  41   b  that leads to the internal space  40   b  is formed at the top end of the nozzle tip  4   b , and an intake and discharge hole  42   b  that leads to the nozzle mounting hole  41   b  through the internal space  40   b  is formed in the distal end plate  43   b  of the nozzle tip  4   b.    
      The nozzle tip  4   b  is designed so that a nozzle  16   b  can be mounted in the internal space  40   b  through the nozzle mounting hole  41   b  (see  FIG. 12 ), and the intake and discharge forces produced by the nozzle  16   b  can be transmitted through the internal space  40   b  and the intake and discharge hole  42   b  to the outside of the nozzle tip  4   b.    
      As shown in  FIG. 7 , a filter  6   b  is provided in the internal space  40   b  of the nozzle tip  4   b . As shown in  FIG. 7 , the filter  6   b  is provided so as to be located near the intake and discharge hole  42   b , which prevents any spray of liquid from getting into the internal space  40   b  from the intake and discharge hole  42   b , and thereby preventing the contamination of the internal space  40   b.    
      The outside diameter of the second side plate  45   b  of the nozzle tip  4   b  is substantially the same as the inside diameter of the second side plate  34   b  of the cover member  3   b , and when the nozzle tip  4   b  is mounted in the nozzle tip fitting space  30   b  of the cover member  3   b , the outer peripheral surface of the second side plate  45   b  of the nozzle tip  4   b  fits snugly against the inner peripheral surface of the second side plate  34   b  of the cover member  3   b  (see  FIG. 8 ( ii )).  
      As shown in  FIG. 7 , the concave component  49   b  is provided on the outer peripheral surface of the second side plate  45   b  of the nozzle tip  4   b , and the concave component  49   b  mates with the convex component  37   b  provided on the inner peripheral surface of the second side plate  34   b  of the cover member  3   b  (see  FIG. 8 ( ii )).  
      When the nozzle tip  4   b  is mounted in the nozzle tip fitting space  30   b  of the cover member  3   b , as shown in  FIG. 8 ( ii ), the inner peripheral surface of the second side plate  34   b  of the cover member  3   b  fits snugly against the outer peripheral surface of the second side plate  45   b  of the nozzle tip  4   b , the nozzle tip fitting hole  31   b  is sealed off by the distal end plate  43   b  and the first and second side plates  44   b  and  45   b  of the nozzle tip  4   b , and the nozzle tip fitting space  30   b  is closed off. The term “closed off” as used here means that there are no gaps, slits, or the like that would hinder the transmission of the intake force or discharge force of the nozzle  16   b  to the nozzle tip fitting space  30   b , and a state in which the nozzle tip fitting space  30   b  leads to the intake and discharge hole  42   b  of the nozzle tip  4   b  is encompassed by “closed off.” 
      Also, when the nozzle tip  4   b  is mounted in the nozzle tip fitting space  30   b  of the cover member  3   b , as shown in  FIG. 8 ( ii ), the convex component  37   b  provided to the second side plate  34   b  of the cover member  3   b  fits into the concave component  49   b  provided to the second side plate  45   b  of the nozzle tip  4   b , so that the nozzle tip  4   b  is fixed to the cover member  3   b , which makes the mounting of the nozzle tip  4   b  in the nozzle tip fitting space  30   b  more secure.  
      Also, when the nozzle tip  4   b  is mounted in the nozzle tip fitting space  30   b  of the cover member  3   b , as shown in  FIG. 8 ( ii ), the abutting surface  38   b  provided to the top end of the second side plate  34   b  of the cover member  3   b  abuts against the third side plate  46   b  of the nozzle tip  4   b , which defines the location of the distal end plate  43   b  of the nozzle tip  4   b  in the nozzle tip fitting space  30   b  (in this embodiment, the distal end plate  43   b  of the nozzle tip  4   b  is defined so as not to come into contact with the bottom plate  32   b  of the cover member  3   b  and seal off the intake and discharge hole  42   b  of the nozzle tip  4   b ), and forms a tightly closed space S 3   b  that leads to the intake and discharge hole  42   b  of the nozzle tip  4   b  between the bottom plate  32   b  of the cover member  3   b  and the distal end plate  43   b  of the nozzle tip  4   b . The tightly closed space S 3   b  has no opening other than the intake and discharge hole  42   b  of the nozzle tip  4   b , so the intake and discharge forces produced by the nozzle  16   b  can be efficiently transmitted from the intake and discharge hole  42   b  of the nozzle tip  4   b  to the tightly closed space S 3   b.    
      When the cover member  3   b  is mounted on the reaction vessel main body  2   b  and the nozzle tip  4   b  is mounted to the cover member  3   b , the tightly closed space S 1   b , as shown in  FIG. 8 ( ii ), has a contact surface with the bottom plate  22   b  of the reaction vessel main body  2   b , and also has a contact surface with the bottom plate  32   b  of the cover member  3   b . Also, as shown in  FIG. 8 ( ii ), the tightly closed space S 3   b  has a contact surface with the bottom plate  32   b  of the cover member  3   b . Therefore, the outside of the reaction vessel  1   b  can be made to communicate tightly closed space S 1   b  and the tightly closed space S 3   b  by forming a through-hole in the bottom plate  22   b  of the reaction vessel main body  2   b  and the bottom plate  32   b  of the cover member  3   b  with a puncture needle provided to the outside of the reaction vessel  1   b  (see  FIG. 12 ( iii )). At this point the bottom plate  32   b  of the cover member  3   b  is opposed against the deepest part of the tightly closed space S 1   b  (the bottom plate  22   b  of the reaction vessel main body  2   b  constituting the surface on which the reaction vessel  1   b  is placed), so a through-hole that communicates between the outside of the reaction vessel  1   b  and the tightly closed space S 1   b  and the tightly closed space S 3   b  can be formed in the bottom plate  22   b  of the reaction vessel main body  2   b  and the bottom plate  32   b  of the cover member  3   b  by a puncture needle (such as a puncture needle  51   b  provided to a puncture vessel  5   b ) provided perpendicular or substantially perpendicular to the surface on which the reaction vessel  1   b  is placed (the lower surface of the bottom plate  22   b  of the reaction vessel main body  2   b ) (see  FIG. 12 ( iii )).  
      As shown in  FIG. 9 , the puncture vessel  5   b  pertaining to this embodiment comprises a main body  50   b  and the puncture needle  51   b . The main body  50   b  has a bottom plate that is quadrangular in plan view, a side plate in the form of an angular cylinder that rises up from the edges of the bottom plate, and a flange provided to the top edges of the side plate. A liquid holding space  501   b  surrounded by the bottom plate and the side plate is formed in the main body  50   b , and an opening  502   b  that leads to the liquid holding space  501   b  is formed at the top end of the main body  50   b.    
      The liquid holding space  501   b  of the puncture vessel  5   b  is designed so that a liquid can be introduced through the opening  502   b , and so that the reaction vessel  1   b  can be accommodated (see  FIG. 12 ( iii )).  
      As shown in  FIG. 9 , the puncture needle  51   b  is provided so as to protrude from the bottom plate of the main body  50   b  into the liquid holding space  501   b  and so as to be substantially perpendicular to the surface on which the reaction vessel  1   b  is placed (the upper surface of the bottom plate of the main body  50   b ). When the reaction vessel  1   b , in which the cover member  3   b  is mounted on the reaction vessel main body  2   b , is placed in the liquid holding space  501   b , a through-hole can be formed in the bottom plate  22   b  of the reaction vessel main body  2   b  and the bottom plate  32   b  of the cover member  3   b  (see  FIG. 12 ( iii )).  
      As shown in  FIG. 9 , the distal end of the puncture needle  51   b  is pointed, and the puncture needle  51   b  is made of stainless steel or another such metal capable of puncturing the plastic, glass, or the like constituting the reaction vessel main body  2   b  and cover member  3   b.    
      As shown in  FIG. 9 , the reaction apparatus  10   b  pertaining to this embodiment comprises a reaction vessel installation part  17   b  in which the reaction vessel  1   b  is installed, a puncture vessel installation part  18   b  in which the puncture vessel  5   b  is installed, a nozzle  16   b  capable of taking up and discharging a liquid, a nozzle transfer part  15   b  that moves the nozzle  16   b  in a specific direction, a first temperature controller  11   b  provided to the reaction vessel installation part  17   b , a second temperature controller  13   b , and a temperature controller mounting and removing part  14   b  that moves the second temperature controller  13   b  in a specific direction.  
      As shown in  FIG. 9 , the reaction vessel installation part  17   b  and the puncture vessel installation part  18   b  are provided on a base  100   b , and a space in which the second temperature controller  13   b  and the nozzle  16   b  can move up, down, left, and right is provided above the base  100   b.    
      As shown in  FIG. 9 , the first temperature controller  11   b  is provided to the reaction vessel installation part  17   b , and the reaction vessel  1   b  is installed on the first temperature controller  11   b.    
      As shown in FIGS.  9  to  11 , the first temperature controller  11   b  comprises a heat-blocking ring  10   b , a heat conductor  111   b , a heat-blocking case  112   b , a thermoelectric semiconductor element  113   b , and a heat sink  114   b.    
      As shown in FIGS.  9  to  11 , a space is formed in the approximate center of the heat-blocking ring  110   b  so that the reaction vessel main body  2   b  can be introduced through an opening at the top, and so that the protrusion of the heat conductor  111   b  can be mounted through an opening at the bottom. The reaction vessel main body  2   b  held in this space is supported by the protrusion of the heat conductor  111   b  mounted in the space. The heat-blocking ring  110   b  is made from a ceramic or other heat-blocking material, and is designed to allow the efficient transfer of heat between the heat conductor  111   b  and the reaction vessel main body  2   b.    
      Also, as shown in FIGS.  9  to  11 , an optical fiber mounting hole  115   b  that communicates with the space in which the reaction vessel main body  2   b  is held is provided to the heat-blocking ring  110   b , and optical fibers can be disposed around the first side plate  23   b  (the outer peripheral surface of the first side plate  23   b ) of the reaction vessel main body  2   b  supported by the protrusion of the heat conductor  111   b  by mounting optical fibers in the optical fiber mounting hole  115   b . A plurality of the optical fiber mounting holes  115   b  are provided to the heat-blocking ring  10   b , and optical fibers connected to a light source (not shown) and optical fibers connected to a fluorescent light detector (not shown) are mounted in these optical fiber mounting holes  115   b , so that the excitation light emitted from the light source and through the optical fibers can irradiate the reaction solution held in the tightly closed space S 1   b  through the first side plate  23   b  of the reaction vessel main body  2   b , and the fluorescent light that is emitted from the reaction solution held in the tightly closed space S 1   b  and that is transmitted through the first side plate  23   b  of the reaction vessel main body  2   b  to the outside of the reaction vessel main body  2   b  can be received through the optical fibers and detected by the fluorescent light detector.  
      As shown in FIGS.  9  to  11 , the heat conductor  111   b  comprises a disk and a protrusion. The protrusion fits into the heat-blocking ring  110   b , while the disk comes into contact with the upper surface of the thermoelectric semiconductor element  113   b  provided on the heat sink  114   b . The heat conductor  111   b  is made of copper or another such metal, so any heat generated by the thermoelectric semiconductor element  113   b  can be efficiently transmitted to the reaction vessel main body  2   b.    
      The thermoelectric semiconductor element  113   b  is a type that can be utilized as a cooling element and/or as a heating element, an example of which is a Peltier element. The thermoelectric semiconductor element  113   b  is connected to a power source (not shown), and when power is supplied from this power source, the heat conductor  111   b  can be heated and/or cooled. As shown in FIGS.  9  to  11 , the lower surface of the thermoelectric semiconductor element  113   b  is in contact with the heat sink  114   b , which has cooling fins, and the thermoelectric semiconductor element  113   b  is forcibly cooled by the heat sink  114   b.    
      As shown in FIGS.  9  to  11 , the heat conductor  111   b  and the thermoelectric semiconductor element  113   b  are held inside the heat-blocking case  112   b , which is made of a ceramic or other heat-blocking material, so the heat conductor  111   b  can be efficiently cooled and/or heated by the thermoelectric semiconductor element  113   b.    
      The first temperature controller  11   b  transmits the heat applied to the heat conductor  111   b  by the thermoelectric semiconductor element  113   b  through the contact surface between the lower surface of the reaction vessel main body  2   b  and the protrusion of the heat conductor  111   b  to the reaction vessel main body  2   b , so that the temperature of the reaction solution held in the tightly closed space S 1   b  can be controlled by the movement of heat through the bottom plate  22   b  of the reaction vessel main body  2   b.    
      As shown in FIGS.  9  to  11 , the second temperature controller  13   b  comprises a heat-blocking ring  130   b , a heat conductor  131   b , a heat-blocking case  132   b , a thermoelectric semiconductor element  133   b , a heat sink  134   b , and an arm attachment component  135   b  to which is attached an extending arm  142   b  of the temperature controller mounting and removing part  14   b.    
      As shown in FIGS.  9  to  11 , a space is formed in the approximate center of the heat-blocking ring  130   b  in which the protrusion of the heat conductor  131   b  can be inserted through the opening on top, and the cover member  3   b  can be inserted through the opening on the bottom. The heat-blocking ring  130   b  is made from a ceramic or other heat-blocking material, and is designed to allow the efficient transfer of heat between the heat conductor  131   b  and the cover member  3   b.    
      As shown in FIGS.  9  to  11 , the heat conductor  131   b  comprises a disk and a protrusion. The protrusion is inserted into the heat-blocking ring  130   b , while the disk comes into contact with the lower surface of the thermoelectric semiconductor element  133   b . The protrusion of the heat conductor  131   b  is formed so that it can be mounted in the nozzle tip fitting space  30   b  of the cover member  3   b , and the protrusion of the heat conductor  131   b  mounted in the nozzle tip fitting space  30   b  is in contact with the bottom plate  32   b , the first side plate  33   b , and the second side plate  34   b  of the cover member  3   b . The outside diameter of the protrusion of the heat conductor  131   b  is smaller than the inside diameter of the heat-blocking ring  130   b , and when the protrusion of the heat conductor  131   b  is inserted into the heat-blocking ring  130   b , a gap that leads to the bottom opening in the heat-blocking ring  130   b  is formed between the outer peripheral surface of the protrusion of the heat conductor  131   b  and the inner peripheral surface of the heat-blocking ring  130   b . The cover member  3   b  can be inserted into this gap, and even when the protrusion of the heat conductor  131   b  has been inserted into the heat-blocking ring  130   b , the protrusion of the heat conductor  131   b  can still be mounted in the nozzle tip fitting space  30   b  of the cover member  3   b . The heat conductor  131   b  is made of copper or another such metal, so any heat generated by the thermoelectric semiconductor element  133   b  can be efficiently transmitted to the cover member  3   b.    
      The thermoelectric semiconductor element  133   b  is a type that can be utilized as a cooling element and/or as a heating element, an example of which is a Peltier element. The thermoelectric semiconductor element  133   b  is connected to a power source (not shown), and when power is supplied from this power source, the heat conductor  131   b  can be heated and/or cooled. As shown in FIGS.  9  to  11 , the upper surface of the thermoelectric semiconductor element  133   b  is in contact with the heat sink  134   b , which has cooling fins, and the thermoelectric semiconductor element  131   b  is forcibly cooled by the heat sink  134   b.    
      As shown in FIGS.  9  to  11 , the heat conductor  131   b  and the thermoelectric semiconductor element  133   b  are held inside the heat-blocking case  132   b , which is made of a ceramic or other heat-blocking material, so the heat conductor  131   b  can be efficiently cooled and/or heated by the thermoelectric semiconductor element  133   b.    
      The second temperature controller  13   b  is designed so that when the protrusion of the heat conductor  131   b  is mounted in the nozzle tip fitting space  30   b  of the cover member  3   b , the heat applied to the heat conductor  131   b  by the thermoelectric semiconductor element  133   b  is transmitted through the contact surface between the protrusion of the heat conductor  111   b  and the bottom plate  32   b , the first side plate  33   b , and the second side plate  34   b  of the cover member  3   b , so that the temperature of the reaction solution held in the tightly closed space S 1   b  can be controlled by the movement of heat through the bottom plate  32   b  of the cover member  3   b.    
      As shown in  FIG. 9 , the temperature controller mounting and removing part  14   b  comprises a rail  140   b  provided substantially perpendicular to the upper surface of the base  10   b , a movable component  141   b  that can move along the rail  140   b , and the extending arm  142   b  provided to the movable component  141   b.    
      As shown in  FIG. 9 , the extending arm  142   b  is provided to the movable component  141   b  so as to be able of extend and retract horizontally with respect to the upper surface of the base  10   b . As shown in  FIG. 9 , the second temperature controller  13   b  is attached via the arm attachment component  135   b  to the distal end of the extending arm  142   b , and the second temperature controller  13   b  is moved horizontally with respect to the upper surface of the base  100   b  by the extension or retraction of the extending arm  142   b , and is moved vertically with respect to the upper surface of the base  100   b  by the movement of the movable component  141   b.    
      The temperature controller mounting and removing part  14   b  is designed so that the protrusion of the heat conductor  131   b  can be mounted in the nozzle tip fitting space  30   b  of the cover member  3   b  of the reaction vessel  1   b  placed on the first temperature controller  11   b , or removed from the nozzle tip fitting space  30   b , by moving the second temperature controller  13   b  horizontally or vertically with respect to the upper surface of the base  100   b.    
      The nozzle  16   b  is connected to a liquid intake and discharge apparatus (not shown) and is designed so that a liquid can be taken up and discharged through an intake and discharge hole  160   b . The intake and discharge hole  160   b  leads to the distal end of the nozzle  16   b , and is designed so that intake force and discharge force can be transmitted to the nozzle tip  4   b  mounted at the distal end of the nozzle  16   b  through an O-ring or the like.  
      As shown in  FIG. 9 , the nozzle transfer part  15   b  comprises a rail  150   b  provided horizontally with respect to the upper surface of the base  100   b , a movable component  151   b  that can move along the rail  150   b , and an extending arm  152   b  provided to the movable component  151   b.    
      As shown in  FIG. 9 , the extending arm  152   b  is provided to the movable component  151   b  so as to be able of extend and retract vertically with respect to the upper surface of the base  10   b . As shown in  FIG. 9 , the nozzle  16   b  attached to the distal end of the extending arm  152   b  is moved vertically with respect to the upper surface of the base  100   b  by the extension or retraction of the extending arm  152   b , and is moved horizontally with respect to the upper surface of the base  100   b  by the movement of the movable component  151   b.    
      The nozzle transfer part  15   b  is designed so that the nozzle tip  4   b  mounted on the nozzle  16   b  can be mounted in the nozzle tip fitting space  30   b  of the cover member  3   b  of the reaction vessel  1   b  placed on the first temperature controller  11   b  by moving the nozzle  16   b  horizontally or vertically with respect to the upper surface of the  10   b . Further, the nozzle transfer part  15   b  is designed so that the reaction vessel  1   b  to which the nozzle tip  4   b  is mounted is moved to the puncture vessel installation part  18   b  and introduced through an opening  502   b  into a liquid holding space  501   b  of the puncture vessel  5   b  placed in the puncture vessel installation part  18   b , and a through-hole can be formed in the bottom plate  22   b  of the reaction vessel main body  2   b  and the bottom plate  32   b  of the cover member  3   b  by the puncture needle  51   b  provided to the puncture vessel  5   b.    
      Furthermore, the operation for the temperature controller mounting and removing part  14   b  and the operation for the nozzle transfer part  15   b  are controlled so as not to interfere with each other.  
      The operation for the reaction apparatus  10   b  will be described by using as an example a case in which a PCR reaction solution is held in the reaction vessel  1   b  and a PCR is conducted.  
      After the PCR reaction solution has been introduced through the opening  21   b  into the reaction chamber  20   b  of the reaction vessel main body  2   b , the cover member  3   b  is placed over the reaction vessel main body  2   b . At this point the convex component  27   b  of the reaction vessel main body  2   b  fits into the concave component  36   b  of the cover member  3   b , and the cover member  3   b  is fixed to the reaction vessel main body  2   b  (see  FIG. 8 ( i )). Also, the tightly closed space S 1   b  and tightly closed space S 2   b  are formed inside the reaction chamber  20   b  when the cover member  3   b  is put in place, so that the PCR reaction solution is held in the tightly closed space S 1   b , and any surplus PCR reaction solution that will not be held in the tightly closed space S 1   b  is held in the tightly closed space S 2   b  (see  FIG. 8 ( i )). The reaction vessel  1   b  in this state is placed in the first temperature controller  11   b  provided to the reaction vessel installation part  17   b  (see  FIGS. 9 and 11 ).  
      The reaction apparatus  10   b  performs an operation in which the second temperature controller  13   b  is moved by the temperature controller mounting and removing part  14   b  to the reaction vessel  1   b  placed in the first temperature controller  11   b , and the protrusion of the heat conductor  131   b  of the second temperature controller  13   b  is mounted in the nozzle tip fitting space  30   b  of the cover member  3   b  (see  FIGS. 9 and 11 ).  
      The reaction apparatus  10   b  also performs an operation in which, after the protrusion of the heat conductor  131   b  has been mounted in the nozzle tip fitting space  30   b , the temperature of the PCR reaction solution held in the tightly closed space S 1   b  is controlled by the first temperature controller  11   b  and the second temperature controller  13   b . As a result, the PCR proceeds in the PCR reaction solution held in the tightly closed space S 1   b , and PCR amplified fragments  7   b  are produced as the reaction product in the PCR reaction solution (see  FIG. 12 ( i )).  
      At this point the PCR reaction solution is held in the form of a thin layer in the tightly closed space S 1   b , so the ratio of surface area to volume is greater, and furthermore nearly all of this surface area is accounted for by the upper and lower surfaces of the thin layer, that is, by the lower surface (contact surface of the cover member) of the bottom plate  32   b  of the cover member  3   b  and the upper surface (opposing surface of the reaction chamber) of the bottom plate  22   b  of the reaction vessel main body  2   b . Therefore, the temperature of the PCR reaction solution held in the tightly closed space S 1   b  can be rapidly controlled by the movement of heat through the bottom plate  32   b  of the cover member  3   b  and by the movement of heat through the bottom plate  22   b  of the reaction vessel main body  2   b , which means that the PCR takes less time.  
      Also, the progress of the PCR (such as whether or not the target nucleic acids have been amplified by PCR, or the amount of PCR amplification product) can be monitored in real time by irradiating the PCR reaction solution held in the tightly closed space S 1   b  with excitation light emitted from the light source, and receiving the fluorescent light emitted from the PCR reaction solution held in the tightly closed space S 1   b  and detecting this light with a fluorescent light detector, through optical fibers mounted in the optical fiber mounting hole  115   b  of the heat-blocking ring  10   b.    
      The reaction apparatus  10   b  also performs an operation in which, after completion of the PCR, the second temperature controller  13   b  is moved by the temperature controller mounting and removing part  14   b , and the protrusion of the heat conductor  131   b  of the second temperature controller  13   b  is removed from the nozzle tip fitting space  30   b  of the cover member  3   b  (see  FIG. 9 ).  
      The reaction apparatus  10   b  also performs an operation in which, after the protrusion of the heat conductor  131   b  has been removed from the nozzle tip fitting space  30   b , the nozzle  16   b  is moved by the nozzle transfer part  15   b  to above the reaction vessel  1   b  placed in the first temperature controller  11   b , and the nozzle tip  4   b  mounted on the nozzle  16   b  is mounted in the nozzle tip fitting space  30   b  through the nozzle tip fitting hole  31   b  (see  FIG. 12 ( i ) and ( ii )). At this point the convex component  37   b  of the cover member  3   b  fits into the concave component  49   b  of the nozzle tip  4   b , fixing the nozzle tip  4   b  to the cover member  3   b . Also, the tightly closed space S 3   b  that leads to the intake and discharge hole  42   b  of the nozzle tip  4   b  is formed inside the nozzle tip fitting space  30   b  by mounting the nozzle tip  4   b  in the nozzle tip fitting space  30   b.    
      The reaction apparatus  10   b  also performs an operation in which, after the nozzle tip  4   b  mounted on the nozzle  16   b  has been mounted in the nozzle tip fitting space  30   b , the nozzle  16   b  is moved by the nozzle transfer part  15   b , and the reaction vessel  1   b  in which is placed the nozzle tip  4   b  mounted on the nozzle  16   b  is moved to above the puncture vessel installation part  18   b  (see  FIG. 9 ). Since the cover member  3   b  is fixed to the reaction vessel main body  2   b , and the nozzle tip  4   b  is fixed to the cover member  3   b , the cover member  3   b  does not come out of the reaction vessel main body  2   b  during movement, nor does the nozzle tip  4   b  come out of the cover member  3   b.    
      The reaction apparatus  10   b  also performs an operation in which, after the reaction vessel  1   b  has been moved to above the puncture vessel installation part  18   b , the extending arm  152   b  is extended, the reaction vessel  1   b  is introduced through the opening  502   b  into the liquid holding space  501   b  of the puncture vessel  5   b  placed in the puncture vessel installation part  18   b  (at this point, the lower surface of the bottom plate  22   b  of the reaction vessel main body  2   b  is pressed against the puncture needle  51   b  provided to the puncture vessel  5   b ), and a through-hole that communicates between the liquid holding space  501   b  of the puncture vessel  5   b , the tightly closed space S 1   b  of the reaction vessel  1   b , and the nozzle tip fitting space  30   b  is formed in the bottom plate  22   b  of the reaction vessel main body  2   b  and the bottom plate  32   b  of the cover member  3   b  by the puncture needle  51   b  provided to the puncture vessel  5   b  (see  FIG. 12 ( c )). At this point the puncture needle  51   b  punctures the bottom plate  22   b  of the reaction vessel main body  2   b , forming a through-hole that communicates between the liquid holding space  501   b  of the puncture vessel  5   b  and the tightly closed space S 1   b  of the reaction vessel  1   b , and then punctures the bottom plate  32   b  of the cover member  3   b , forming a through-hole that communicates between the tightly closed space S 1   b  of the reaction vessel  1   b  and the tightly closed space S 3   b  inside the nozzle tip fitting space  30   b.    
      After the reaction vessel  1   b  has been punctured by the puncture needle  51   b , the liquid holding space  501   b  of the puncture vessel  5   b  communicates with the tightly closed space S 1   b  of the reaction vessel  1   b  through the through-hole formed in the bottom plate  22   b  of the reaction vessel main body  2   b , and the tightly closed space S 1   b  of the reaction vessel  1   b  communicates with the tightly closed space S 3   b  inside the nozzle tip fitting space  30   b  through the through-hole formed in the bottom plate  32   b  of the cover member  3   b , and since the tightly closed space S 3   b  inside the nozzle tip fitting space  30   b  leads to the intake and discharge hole  42   b  of the nozzle tip  4   b , the intake force and discharge force produced by the nozzle  16   b  can be transmitted to the liquid holding space  501   b  of the puncture vessel  5   b.    
      The reaction apparatus  10   b  also performs an operation in which, after the puncture by the puncture needle  51   b , intake and discharge by the nozzle  16   b  are commenced, and the extract  8   b  (such as a buffer) held in the liquid holding space  501   b  of the puncture vessel  5   b  is taken up and discharged through the above-mentioned through-holes, so that the PCR amplified fragments  7   b  contained in the PCR reaction solution in the tightly closed space S 1   b  of the reaction vessel  1   b  are extracted into the extract  8   b  (see  FIG. 12 ( iii )). At this point, when the intake and discharge by the nozzle  16   b  are commenced, the extract  8   b  held in the liquid holding space  501   b  of the puncture vessel  5   b  flows into the tightly closed space S 1   b  along with intake by the nozzle  16   b , and flows out of the tightly closed space S 1   b  along with discharge by the nozzle  16   b . As the intake and discharge of the nozzle  16   b  are repeated over and over, this inflow of the extract  8   b  to the tightly closed space S 1   b  and outflow from the tightly closed space S 1   b  is also repeated, so that the PCR amplified fragments  7   b  contained in the PCR reaction solution in the tightly closed space S 1   b  of the reaction vessel  1   b  are extracted into the extract  8   b  held in the liquid holding space  501   b  of the puncture vessel  5   b.    
      After a PCR has thus been conducted when the cover member  3   b  covering the reaction vessel main body  2   b , the PCR amplified fragments  7   b  contained in the PCR reaction solution in the tightly closed space S 1   b  of the reaction vessel  1   b  can be acquired without removing the cover member  3   b  from the reaction vessel main body  2   b.    
      The second embodiment described above was given in order to facilitate an understanding of the present invention, and does not limit the present invention in any way. Therefore, the various elements disclosed in the second embodiment should be construed as encompassing all design modifications, equivalents, etc., within the technological scope of the present invention.  
      When the reaction chamber  20   b  is closed off by covering the reaction vessel main body  2   b  with the cover member  3   b , the inner peripheral surface of the third side plate  25   b  of the reaction vessel main body  2   b  does not have to be in direct contact with the outer peripheral surface of the second side plate  34   b  of the cover member  3   b , and may instead have a member capable of maintaining a seal, such as an O-ring, interposed between these members. Similarly, when the nozzle tip fitting space  30   b  is closed by mounting the nozzle tip  4   b  in the nozzle tip fitting space  30   b  of the cover member  3   b , the inner peripheral surface of the second side plate  34   b  of the cover member  3   b  does not have to be in direct contact with the outer peripheral surface of the second side plate  45   b  of the nozzle tip  4   b , and may instead have a member capable of maintaining a seal, such as an O-ring, interposed between these members. Here, a gap, slit, or the like that communicates with the inside and outside of the reaction chamber  20   b  or of the nozzle tip fitting space  30   b  may be formed ahead of time in the O-ring or other member, so that any air inside the reaction chamber  20   b  or the nozzle tip fitting space  30   b  can be discharged to the outside when the reaction vessel main body  2   b  is covered with the cover member  3   b  or when the nozzle tip  4   b  is mounted in the nozzle tip fitting space  30   b . Also, a gap, slit, or the like that communicates with the inside and outside of the reaction chamber  20   b  or of the nozzle tip fitting space  30   b  may be formed ahead of time in the inner peripheral surface of the third side plate  25   b  of the reaction vessel main body  2   b  or the outer peripheral surface of the second side plate  34   b  of the cover member  3   b , or in the inner peripheral surface of the second side plate  34   b  of the cover member  3   b  or the outer peripheral surface of the second side plate  45   b  of the nozzle tip  4   b.    
     INDUSTRIAL APPLICABILITY  
      The present invention provides a reaction vessel, a reaction apparatus, and a method with which a reaction can be automated without requiring centrifugation when a reaction solution is held in a reaction chamber, the temperature of the reaction solution held in the reaction chamber can be rapidly controlled, the reaction can proceed even when just a tiny amount of reaction solution is held in the reaction chamber, and the reaction occurring in the reaction chamber can be monitored in real time (that is, instantly during the course of the reaction).  
      The present invention also provides a reaction vessel, a reaction apparatus, and a method with which, after a reaction has been conducted with the cover member covering the reaction vessel main body, the reaction product contained in the reaction solution inside the reaction vessel can be acquired without removing the cover member from the reaction vessel main body.  
      With the reaction vessel, reaction apparatus, and method of the present invention, a series of operations comprising the preparation of samples containing target nucleic acids (such as extraction of nucleic acids from cells), amplification of these target nucleic acids by PCR, monitoring of the progress of the PCR (such as whether or not the target nucleic acids have been amplified, or the amount of PCR amplification product), and acquiring PCR amplified fragments can be automated, making it possible for numerous specimens to be processed in parallel and efficiently.