METHOD FOR CONTROLLING LIQUID TRANSPORT IN A FLOW PATH OF A BIOMOLECULAR ANALYST USING A COMPUTER AND BIOMOLECULAR CLEANING SYSTEM

Abstract

To prevent air from flowing beyond a cleaning membrane, even without an additional flow path structure, the present disclosure proposes a method for controlling the liquid transport in a flow path of a biomolecule analyzer by means of a computer. The biomolecule analyzer comprises a first chamber storing a first liquid, a second chamber storing a second liquid, a membrane chamber with a cleaning membrane, and a waste liquid chamber.The method comprises controlling the transport of the second liquid until it has flowed at least beyond a merging point between a first flow path leading from the first chamber to the waste liquid chamber and a second flow path originating from the second chamber, and expelling a fluid different from the first and second liquids from the second flow path by the computer, controlling the transport of the first liquid from the first chamber via the membrane chamber to the waste liquid chamber by the computer, and controlling the transport of the second liquid in the second chamber from the second chamber to the membrane chamber by the computer.

Description

Technical area

[0001] The present disclosure relates to a method for controlling the fluid transport in a flow path of a biomolecule analyzer by means of a computer and a biomolecule purification system. Technical background

[0002] When genes are analyzed, for example, sample lysis and nucleic acid purification and amplification are performed as pretreatments, and then the amplified product is detected. This process carries the risk of contamination and requires complex sample preparation. Therefore, the traditional procedure involved sending a sample to a laboratory-style experimental setup, where a technician with specialized knowledge and techniques performed sample preparation and measurement to analyze the data. However, this approach has the drawbacks of time-consuming sample transport and high equipment and laboratory costs to maintain the setup. Furthermore, if batch processing is implemented to increase efficiency, it is difficult to interrupt the process for an urgent sample.

[0003] In recent years, StoA (Sample-to-Answer) systems have emerged in various fields. These systems operate fully automatically from receiving a sample to performing a measurement and data acquisition. The StoA system utilizes a flow path chip that integrates a chamber, a flow path, and a reagent. The introduction of the flow path chip provides the following advantages: (i) measurement can be easily performed by a non-expert, (ii) data can be acquired in a short time, (iii) a highly portable device can be developed, (iv) variation in a procedure is reduced, and (v) reagent storage is facilitated.

[0004] The application areas of the StoA flow path chip include forensic medicine, in vitro diagnostics, animal and plant species identification, biodefense, medicine, biotechnology, life sciences, defense, public health, and agriculture, among other potential applications. If genetic analysis is performed on a StoA flow path chip, it is desirable that part or all of the flow path directly contacting the sample be interchangeable for each measurement to prevent sample mixing between analyses. To reduce the cost of the disposable chip, measures such as an easy-to-manufacture design and the use of inexpensive materials are taken.

[0005] However, with such a cost-effective chip, the compressive strength is limited by a valve, the chip bond strength, and similar factors. In the case of PTL 1, for example, a flow path chip has a simple structure in which two thermoplastic resins are bonded together. The compressive strength of this chip is limited by a valve and is described as being 68 kPa. For example, in the case of PTL 2, the compressive strength of the flow path chip is described as being 124 kPa. In contrast, in large-scale flow path systems, such as those used in liquid chromatography, the pressure usable for solution transport is more than a few MPa. In the case of spin columns, which are widely used in nucleic acid purification, pressures up to 500 kPa can be applied.As described above, the pressure that can be used when sample processing is performed in the StoA flow path chip is usually lower than that in a benchtop system.

[0006] In the StoA system, the solution transport must be automated under limited space and pressure. Accordingly, it is advantageous if there are no air bubbles in the flow path. This is because there are concerns that the solution transport will be incomplete due to air trapped in the flow path or that an unexpected process will be caused. To solve the problem caused by the presence of air in such a flow path, according to NPL 1, a material through which air escapes is used for the flow path. According to NPL 2 and PTL 3, an air removal structure is installed in a flow path. This problem can also be solved by compressing or moving the air under high pressure. ZitatlistePatentliteratur PTL 1: US-Patent Nr. 10233491 PTL 2: US-Patent Nr. 9354199 PTL 3: japanisches Patent Nr. 6613212 Nicht-Patentliteratur NPL 1: „PDMS membranes with tunable gas permeability for microfluidic applications - RSC Advances (RSC Publishing) DOI:10.1039 / C4RA1293B“, RSC Adv., 2014, 4, 61415 NPL 2: „Integrated Microfluidic System for Rapid Forensic DNA Analysis: Sample Collection to DNA Profile / Analytical Chemistry (acs.org)“, Anal. Chem. 2010, 82, 16, 6991-6999 Kurzfassung der ErfindungTechnisches Problem

[0007] In the purification and recovery of nucleic acids using a purification membrane, for example made of silica, high recovery efficiency can be achieved with a simple flow path structure. However, if this purification process is carried out in a flow path with a confined space, allowing air to escape, the following problems arise. If air enters the chamber housing the membrane, it must pass through the membrane chamber. For example, after a lysate has passed through a membrane, if air passes through the membrane while it is wetted with the lysate, the applied pressure must exceed a Laplace pressure PL defined by the following expression (1). [Math. 1] PL=4γcosθd

[0008] Here, θ, d, and γ represent the contact angle between the liquid wetting the film and the film, the pore diameter of the membrane, and the surface tension of the liquid, respectively. If a fine-pored silica membrane is used for cleaning, the Laplace pressure is assumed to be significant because d is small.

[0009] In this respect, providing a structure for venting air, as in the techniques disclosed in PTL 3, NPL 1 and NPL 2, can prevent air from passing through the membrane.

[0010] However, if this approach is used, an additional structure must be provided in the flow path. There are limitations regarding the materials that can be used for this additional structure, and concerns about the increased cost and size of the chip.

[0011] Furthermore, in the case of PTL 1, the purification membrane installed in the purification chamber does not cover the entire surface of the flow path. Because the air overflows beyond the side of the purification membrane, the problem described above does not occur. However, because the proportion of lysate in contact with the membrane decreases, there are concerns that the recovery rate will decrease.

[0012] In view of this situation, the present disclosure proposes a technique to prevent a fluid (air, nitrogen and other gases) from overflowing a cleaning membrane, even without an additional flow path structure. Solution to the problem

[0013] To solve the above problems, the present disclosure provides, for example, a method for controlling fluid transport in a flow path of a biomolecule analyzer by a computer. The biomolecule analyzer includes a first chamber storing a first fluid, a second chamber storing a second fluid, a membrane chamber including a purification membrane, and a waste fluid chamber.The method comprises controlling the transport of the second liquid until it has flowed at least beyond a merging point between a first flow path leading from the first chamber to the waste liquid chamber and a second flow path originating from the second chamber, and expelling a fluid different from the first and second liquids from the second flow path by the computer, controlling the transport of the first liquid from the first chamber via the membrane chamber to the waste liquid chamber by the computer, and controlling the transport of the second liquid in the second chamber from the second chamber to the membrane chamber by the computer.

[0014] Further features of the present disclosure will become clear with reference to the present description and the accompanying drawings. Aspects of the present disclosure are achieved and realized through elements, combinations of various elements, and aspects of the following detailed description and the accompanying claims. The present description serves only as an example and is not intended to limit the scope of the claims or application examples of the present disclosure in any way. Advantageous effects of the invention

[0015] The technology described in this disclosure makes it possible to prevent air from flowing beyond the cleaning membrane even without an additional flow path structure. Brief description of the drawings

[0016] They show: Fig. 1 a diagram of a configuration example of a biomolecule analyzer 100 according to the present embodiment, Fig. 2 a diagram of a derived form of part of the biomolecule analyzer 100 and a flow path chip 114, Fig. 3 a diagram of an example procedure for performing a biological analysis using the Biomolecule Analyzer 100, Fig. 4 a diagram of an example of the internal configuration of a computer 115, Fig. 5 is a diagram of a configuration example of a purification system 301 having a main part of a flow path chip 114 according to a first example, which is a main part of the flow path chip 114 upon completion or execution of a sample lysing procedure 202, Fig. 6 a diagram of cleaning processes I to V according to the first example, Fig. 7 a flowchart that corresponds to the one in Fig. The process shown in section 6 corresponds to Fig. 8. A diagram to explain a cleaning process according to a comparative example. Fig. 9 a flowchart of a cleaning process according to Fig. 8, Fig. 10 a schematic view of the position of a solution during transport and a diagram of a pressure change measured according to the experiment of the first example, Fig. 11 is a diagram of a configuration example of a purification system 400 having a main part of a flow path chip 114 according to a second example, which is a main part of the flow path chip 114 upon completion or execution of a sample lysing procedure 202, Fig. 12 a diagram to explain the cleaning processes I to V according to the second example, Fig. 13 a diagram of a configuration example of a cleaning system 500 with a main part of a flow path chip 114 according to a third example, wherein the main part of the flow path chip 114 is at the completion or execution of a sample lysis procedure 202, Fig. 14 a diagram to explain the cleaning processes I to V according to the third example, Fig. 15 a flow chart corresponding to Fig. 14 corresponds to the cleaning process shown, Fig. 16 a diagram of a configuration example of a cleaning system 600 with a main part of a flow path chip 114 according to a fourth example, wherein the main part of the flow path chip 114 is at the completion or execution of a sample lysis procedure 202, Fig. 17 a diagram to explain the cleaning processes I to V according to the fourth example, Fig. 18 a diagram explaining the cleaning processes I to V according to a fifth example and Fig. 19 a diagram explaining the cleaning processes I to V according to a sixth example. Description of embodiments

[0017] The present embodiment proposes a technique for installing a purification membrane in a flow path chip, where structural robustness is not a requirement, by reducing the pressure required for fluid transport in the flow path chip in each example. First, each feature of the biomolecule analyzer according to the present embodiment is described, and then the respective examples are described. In the drawings accompanying this application, functionally identical elements may be designated with the same numbers. It should be noted that the accompanying drawings are for the purpose of understanding the present disclosure and should not be used to interpret the present disclosure restrictively, even though the accompanying drawings show specific embodiments and implementation examples that are consistent with the basic ideas of the present disclosure.Furthermore, the present embodiment has been described in sufficient detail to enable those skilled in the art to implement the present disclosure. However, it should be understood that other implementations and modes are possible, and changes can be made to the configurations and structures, and various elements can be replaced, without deviating from the scope and the fundamental idea of ​​the technical concept presented in the present disclosure. Therefore, the following description should not be interpreted as being limited to this.

[0018] In the embodiments of the present disclosure, the computer-controlled operation can be implemented by software running on a general-purpose computer, or it can be implemented by dedicated hardware or a combination of software and hardware. (1) Features of the biomolecule analyzer<Strömungsweg-Chip>

[0019] According to the present embodiment, "flow path chip (or simply chip)" refers to a disposable or reusable cartridge comprising a reagent, a chamber, and a flow path. The flow path chip may include a solution transport current source. Some or all of the reagents may be contained within the chip. A portion of the chamber may incorporate a temperature control function, a molecule trapping function, a detection function, and a voltage application function.

[0020] The material of the flow path chip is not particularly restricted, as long as it is a material commonly used in the technical field. For example, polypropylene, a cyclic olefin polymer (COP), a cyclic olefin copolymer (COC), polycarbonate, polyethylene terephthalate, or polyurethane are preferably used as materials with a low DNA adsorption capacity. It is also desirable to reduce the adsorption capacity by modifying the surface so that it is negatively charged.Examples of other materials include metals such as gold, silver, copper, aluminum, tungsten, molybdenum, chromium, platinum, titanium, and nickel; alloys such as stainless steel, Hastelloy, Inconel, Monel, and duralumin; silicon; glass materials such as glass, fused silica, molten quartz, synthetic quartz, aluminum oxide, sapphire, ceramics, forsterite, and photosensitive glass; plastics such as polyester resins, polystyrene, polyethylene resins, ABS resins (acrylonitrile butadiene styrene resins), dimethyl polysiloxane (PDMS), nylon, acrylic resins, fluorocarbon resins, polycarbonate resins, polyurethane resins, methylpentene resins, phenolic resins, melamine resins, epoxy resins, and vinyl chloride resins; agarose, dextran, cellulose, polyvinyl alcohol, nitrocellulose, chitin, chitosan, or a combination thereof.

[0021] If the cross-sectional area of ​​the flow path is too large, there are concerns about liquid residue or sample loss. In addition, if the cross-sectional area is too large, it is difficult to apply pressure. On the other hand, if the cross-sectional area is too small, there is the problem that high pressure is required to transport the solution or the time required for solution transport becomes long. Therefore, the cross-sectional area of ​​the flow path can be in the range of 1 µm. 2 up to 314 mm 2 The cross-sectional area can be in the range of 400 µm² in a preferred configuration. 2 up to 100 mm 2 The cross-sectional area can be in the range of 0.01 mm² in an even more preferred configuration. 2 up to 10 mm 2 lay.

[0022] If the flow path length is too short, it becomes difficult to incorporate elements such as valves. If the flow path length is too long, there is a risk of the chip size increasing. Therefore, the length of the flow path connecting the chip configurations can range from 1 µm to 100 cm. A more preferred range is 1 mm to 50 cm. An even more preferred range is 5 mm to 30 cm. <kammer>

[0023] The chamber is a space in which a liquid or solid can be stored, its diameter being equal to or greater than that of the flow path. The reagent can be stored in the chamber, and PCR, lysis, purification, or similar processes can be performed in the chamber.

[0024] For example, the chamber volume is 0.01 µL to 10 L. If the volume is larger than 10 L, the device becomes difficult to carry. In a more preferred example, the chamber volume may be 0.1 µL to 2 L. Note that not all chambers need to be installed in the chip and may also be provided in the device main body or another independent chip.

[0025] If the chamber is installed within the chip, its volume can range from 0.01 µl to 50 ml. If the volume exceeds 50 ml, the chip becomes large and difficult to store.

[0026] The chip stores one or more reagents in one or more reagent storage chambers. If these reagents are mixed at an unintended time, functionality will be compromised and other unexpected results may occur. Therefore, it is desirable that these reagents be separated until immediately before use by a separation mechanism including a valve, a film, air, a flow path thin enough to prevent spontaneous mixing, or a combination thereof. By isolating the reagent from the outside air, long-term storage and portability of the chip can be realized.

[0027] Similarly, if the reagent is stored outside the chip (if the chamber is attached outside the flow path chip), it is desirable to store it in a state isolated from outside air and separated from other components of the cleaning system by a valve, film, air, or the like. <reinigungssystem>

[0028] According to the present embodiment, “cleaning system” means a system in which a cleaning membrane provided in the flow path chip and a membrane chamber receiving the cleaning membrane capture biomolecules contained in the lysate stored in the lysis chamber, and the lysate is subsequently washed with a wash buffer stored in a wash buffer chamber.

[0029] The lysis chamber and the wash buffer chamber may be provided outside the flow path chip, or one or both of them may be provided in the chip.

[0030] The purification system can be designed to selectively capture specific biomolecules. For example, DNA can be selectively extracted from a liquid containing proteins, DNA, ions, and the like. The purification system can be a purification system for obtaining a molecule that contains RNA, a biopolymer (nucleic acids, proteins, lipids, polysaccharides), a biomonomer (amino acids, lipids, sugars, nucleobases), and a derivative thereof in its structure. A system for obtaining multiple types of these molecules can be used. An example is a system capable of obtaining DNA and RNA, which uses a silica membrane as the purification membrane. <probentyp>

[0031] The sample on which the purification system according to the present embodiment is to act is not particularly restricted, as long as it originates from an organism. The organism from which the sample is derived is not particularly restricted, and a sample can be used that originates from an organism of the type of a vertebrate (for example, mammals, birds, reptiles, fish, amphibians, and the like), an invertebrate (for example, insects, nematodes, crustaceans, and the like), a plant, a protist, a fungus, a bacterium, or a virus.

[0032] When collecting a sample, a swab, filter paper, cloth, or similar material can be used as a carrier. The sample can be introduced into the purification system along with the carrier. <lysat>

[0033] When the sample is transported to the membrane chamber, it must be in a form that allows it to flow through the flow path. If the sample is in a solid state (for example, a swab sample), it is therefore preferred to dissolve or suspend the solid sample in a lysis buffer to obtain a liquid lysate. The sample need not be completely dissolved. Any portion that is solid or has a high viscosity after lysis may remain in the lysis chamber. If the sample is gaseous (for example, in the form of air, exhaled air, and the like), it is preferred to obtain a liquid sample by suspending cells contained in the gaseous sample in a solvent. Preparation methods for converting a sample into a lysate are common in the field and readily understood by those skilled in the art. For example, the lysis buffer may contain a chlorinated material such as calcium hypochlorite.In another example, the substance may contain enzymatic activities such as DNAase, RNAase, protease, and the like. The lysis buffer may, if required, contain a substance that facilitates the release of biomolecules, such as chaotropes, a surfactant, and KOH, or a substance that facilitates the binding of nucleic acids to the purification membrane. The mixture may, if necessary, be subjected to a process such as heating and stirring.

[0034] In the present embodiment, “lysate” means a substance obtained by converting a sample derived from an organism into a liquid having a viscosity of 100,000 mPa·s or less using a lysis buffer. In a more preferred condition, the lysate may have a viscosity of 10,000 mPa·s or less. In a still more preferred condition, the lysate may have a viscosity of 1,000 mPa·s or less. <Lösungstransportsteuerung>

[0035] A valve can be used to control the transport of the solution. Solution control can be achieved via the flow path resistance.

[0036] According to the present embodiment (in each example), atmospheric pressure is used as the driving force for solution transport. Air pressure, mechanical compression, centrifugal force, or the like can also be used as the driving force for solution transport.

[0037] If the pressure used for transport is too high, it will exceed the pressure resistance of the chip. If the pressure used for transport is too low, the transport time will be excessive, leading to an increase in the measurement time. Therefore, the pressure used for transport can range from 0.1 kPa to 1 MPa. Within a narrower range, the solution can be transported at 0.1 kPa to 500 kPa, and within an even narrower range, at 0.1 kPa to 200 kPa. The transport time can be up to 1 hour per step, 10 minutes within a narrower range, and 5 minutes within an even narrower range. If the volume of liquid to be transported is too large, the time and pressure required for transport will increase, as will the cost of the chip and the reagent.Therefore, the amount of liquid to be transported per reagent can be 1 liter or less, 10 ml or less in a narrower range, and 2 ml or less in an even narrower range. <waschpuffer>

[0038] In the present embodiment, "wash buffer" means a liquid used for washing away substances adhering to the cleaning membrane that are not required for subsequent steps. The wash buffer may not be able to wash away all unnecessary substances and may wash away some or all of the necessary substances.

[0039] According to the preceding expression (1), a washing buffer with the following features can be used in the present embodiment. First, washing buffers with a higher evaporation rate than the lysate are suitable. Additionally, it is desirable that the lysate and the washing buffer are compatible. Furthermore, it is desirable that the contact angle with the cleaning membrane is smaller (the wettability is lower) or that the interfacial tension is lower than that of the lysate. Examples of washing buffers that meet such a requirement are ethanol and isopropanol. A liquid containing at least 10% of these alcohols can also be used as a washing buffer. A solution that does not meet the above conditions can also be used.

[0040] One, two, or more than two wash buffers can be used. Using two or more types of wash buffers can make the washing process more effective. These wash buffers can be stored in one chamber or in two or more chambers. If two or more types of wash buffers are used, air may or may not be present between them. However, if the second and subsequent wash buffers have a higher evaporation rate, lower surface tension, or a smaller contact angle with the cleaning membrane than the first wash buffer, it is desirable to transport them continuously. <reinigungsmembran>

[0041] Examples of this membrane type include a silica membrane. Other examples of the purification membrane may include a solid substrate consisting mainly of cellulose capable of adsorbing DNA, carboxylated particles, and an ion-exchange resin. In particular, a membrane with a hydroxyl group or a silica group on its surface may be used. The membrane can be of any type, as long as it can retain particles measuring 100 µm or more. The thickness of the membrane is preferably 1 µm or more. Because DNA can be retained more effectively if the membrane pores are finer, a membrane capable of retaining particles measuring 10 µm or more, preferably 1 µm or more, and even more preferably 0.1 µm or more, may be used.

[0042] If the volume of the purification membrane is too small, the amount of adsorbable biomolecules is reduced. Conversely, if the volume of the purification membrane is too large, there are concerns that unintended molecular adsorption will occur during the purification step or a subsequent step, or that the transport efficiency of the solution will be impaired. In each of the examples described later, a membrane with an area of ​​12.5 mm² is used. 2 used. For example, a membrane with an area of ​​1 mm² can also be used. 2 up to 314 mm 2 can be used, and the size of the membrane is not limited. <nachweisverfahren>

[0043] In the present purification system, amplification is performed by PCR. After amplification, detection is performed by capillary electrophoresis (CE). Other examples include massively parallel sequencing (MPS), pyrosequencing, Sanger sequencing, nanopore sequencing, chromatography, electrical measurement, spectroscopy, NMR, restriction fragment length polymorphism (RFLP), and the like. <Andere ergänzende Elemente>

[0044] In all drawings used to describe the present embodiment, components with the same function are designated by the same reference numerals, and their repeated description is avoided as far as possible. Each example of the present embodiment is described in detail below with reference to the accompanying drawings. The measuring method, the structure of the device, the type of substance, and the material described in each example are examples illustrating the concept of the present embodiment and do not strictly specify the measuring principle, the material, the dimensions of the device, etc. Specific pressure values ​​described in each example are examples illustrating the concept of the present embodiment and do not strictly specify them.The specific sample types, composition, and amount of liquid in a cleaning kit described in each example are examples for implementing the concept of the present embodiment and do not serve to strictly define the chemical composition and time. The specific types of measurement objects and solutions and their concentrations described in each example are examples for implementing the concept of the present embodiment and do not serve to strictly define the chemical composition. <Konfigurationsbeispiel des Biomolekülanalysators>

[0045] Fig. Figure 1 is a diagram showing a configuration example of a biomolecule analyzer 100 according to the present embodiment. The biomolecule analyzer 100 includes a flow path chip 114 and a computer 115 for performing biomolecule analysis.

[0046] The flow path chip 114 comprises a lysis chamber 101 for introducing and lyseing a collected sample, a membrane chamber 108 containing a purification membrane 102, a reaction chamber 103 for DNA amplification, a waste fluid chamber 107, and an opening 109 that is in fluid communication with the area outside the chip. The arrangement of the chambers in the flow path chip 114 and the flow path connection between the chambers differ in each example. Therefore, the chamber arrangement, the flow path connection, and the process of transporting the lysate and the wash buffer are described later.

[0047] A GF / F fiberglass membrane with a 4 mm diameter cutout, distributed by Whatman, was installed as the cleaning membrane 102. Other membranes can also be used as cleaning membrane 102. The diameter of the cleaning membrane (the longitudinal length in the case of a rectangle) can range from 0.1 mm to 100 mm.

[0048] The solution is transported through opening 109 by applying pressure, and a reagent, amplification product, and the like can be exchanged with the area outside the chip. The pressure can be applied using a pressure generator provided outside the chip. The flow path chip 114 according to Fig. 1 has chambers 104, 105, and 106 for holding a reagent and the like, and a configuration including the membrane chamber 108. Regarding the membrane chamber 108, the volume of a section located near the lysis chamber 101 in the space separated by the membrane is 10 µl, and the volume of a section near the waste liquid chamber 107 is also 10 µl. The lysis buffer chamber 104 stores a lysis buffer 110 for lyseing a sample. The wash buffer chamber 105 stores a wash buffer 111. The reagent chamber 106 stores an eluate or a reagent 112 used for a reaction. The aforementioned functions may be partially integrated into the same chamber. The positional relationship of the chip components described above is not limited to those shown in the diagram. Fig. 1 limited, and the connection of the flow paths can be determined from that in Fig. 1 may be different.

[0049] Fig. Figure 2 is a diagram showing part of the biomolecule analyzer 100 and a derived form of the flow path chip 114. Fig. The lysis buffer chamber 104, the wash buffer chamber 105, and the waste liquid chamber 107 are provided outside the flow path chip 114. Using this form allows for miniaturization benefits and a cost reduction of the flow path chip 114. <Beispiel einer biologischen Analyseprozedur>

[0050] Fig. Figure 3 is a diagram showing an example of a procedure for performing a biological analysis using the Biomolecule Analyzer 100. As shown in Fig. As shown in Figure 3, the biological analysis includes, for example, a sample reception procedure 201, a sample lysis procedure 202, a sample generation procedure 203, a sample amplification procedure 204 and a sample detection procedure 205.

[0051] In the sample reception procedure 201, the lysis buffer 110 is transported from chamber 104 to lysis chamber 101 before and after the sample has been stored in lysis chamber 101 (insertion of the sample into lysis chamber 101).

[0052] Next, in sample lysis procedure 202, lysis is initiated. In sample generation procedure 203, a lysate 113 is sent from lysis chamber 101 to purification membrane 102 to bind DNA to the membrane, and purification is performed. After purification, a step to dry the wash buffer or the like may be included. The eluted DNA is transported to reaction chamber 103.

[0053] In sample amplification procedure 204, the purified DNA is amplified. In sample detection procedure 205, the amplified DNA is measured. Sample detection procedure 205 can be performed in the flow path chip 114. Alternatively, the amplified DNA can be transported to a detection unit outside the flow path chip 114, which is a separately provided measuring unit.

[0054] It should be noted that procedures 201 to 205 can be executed in parallel. Some procedures can be omitted, or other procedures can be included. <Beispiel der internen Konfiguration und Steuervorgang des Computers 115>

[0055] Fig. Figure 4 is a diagram showing an example of the internal configuration of computer 115. Computer 115 has a processor (not shown), a user interface 1151, and a database (storage device) 1152.

[0056] User interface 1151 has an input screen display and an output screen display. User interface 1151 can receive parameters relating to the implementation procedure from Fig. 3. Retrieve parameters such as time, temperature, pressure, flow rate, procedure, and the like for each step performed by the user, and store these parameters in database 1152. Various parameters can be pre-stored in database 1152.

[0057] The computer 115 can open and close the valve of the flow path chip 114, control the temperature, and control the applied pressure and flow rate based on various parameters stored in the database 1152. Furthermore, the computer 115 can automatically perform all functions in Fig. 3. It should be noted that part of the implementation procedures described in Fig. The implementation procedure shown in section 3 can be supported and implemented by the user. (2) Examples [First example]

[0058] A first example will be given with reference to the Fig. 5 to 7. <Konfigurationsbeispiel des Reinigungssystems 301, das einen Hauptteil des Strömungsweg-Chips 114 aufweist>

[0059] Fig. 5 is a diagram showing a configuration example of a purification system 301 including a main part of the flow path chip 114 according to the first example, that is, a main part of the flow path chip 114 upon completion or execution of the sample lysing procedure 202.

[0060] The purification system 301 includes a lysis chamber 101, a wash buffer chamber 105, a waste liquid chamber 107, a membrane chamber 108, valves 302, 303, 304, 305, and 315, flow paths 306, 307, 308, 309, 310, 311, and 312 connecting the chambers, and an orifice 109. The number of orifices 109 may not be 1, and one orifice may be installed for each function. For example, dedicated orifices may be provided for flow paths 311 and 310, respectively.

[0061] Lysis chamber 101 receives the lysate 113. Wash buffer chamber 105 receives the wash buffer 111. A liquid level detection sensor 313 is installed in the waste liquid chamber 107. A liquid level detection sensor 314 is installed in lysis chamber 101. One or more liquid level detection sensors may be installed, or this may not be the case. Additionally, a liquid level detection sensor may be installed in another chamber of lysis chamber 101 or waste liquid chamber 107.

[0062] As in Fig. As shown in Figure 5, the flow path 306, which originates from the outlet of the lysis chamber 101, and the flow path 307, which originates from the outlet of the wash buffer chamber 105, are joined at a junction point 316, forming the flow path 308, which leads to the inlet of the membrane chamber 108. The flow path 309, which originates from the outlet of the membrane chamber 108, leads to an inlet in the lower section (bottom surface section, although this is not necessarily a bottom surface) of the waste liquid chamber 107. Two flow paths 310 and 311 extend from the inlet 109. Flow path 310 leads to an upper section (surface section, although it is not necessarily an upper surface) of the waste liquid chamber 107. Flow path 311 leads to an inlet opening of the washing buffer chamber 105.The flow path 312 leads to an upper section (surface section, although it is not necessarily an upper surface) of the waste liquid chamber 107.

[0063] As in Fig. As shown in Figure 5, valve 302 is located in flow path 307. Valve 303 is located in flow path 311. Valve 304 is located in flow path 310. Valve 305 is located in flow path 306. Valve 315 is located in flow path 312. <reinigungsprozess>

[0064] Fig. Figure 6 is a diagram showing cleaning processes I to V according to the first example. Fig. 7 is a flowchart that corresponds to the one in Fig. The process shown in Figure 6 corresponds to this. The computer (processor) 115 controls the opening and closing of each valve and the transport of the lysate 113 and the wash buffer 111 by applying pressure in each process. (i) Process I

[0065] Process I shows the state during or immediately after completion of lysis in sample lysis procedure 202. Valves 302, 303, 304, 305 and 315 are then closed (step 701). (ii) Process II

[0066] In Process II, valves 302, 303, and 315 are opened (step 702). Wash buffer 111 is withdrawn from wash buffer chamber 105 at a pressure of 30 kPa or less for a time of 30 seconds or less and supplied to membrane chamber 108 (step 703). At this time, the air in flow path 307 moves to flow path 308 or 306, and flow path 307 is filled with wash buffer 111. Expelling the air in flow path 307 in Process II is important to achieve continuous transport without trapping air between lysate 113 and wash buffer 111. (iii) Process III

[0067] In Process III, valves 302, 303, and 315 are closed, and valves 305 and 304 are opened (step 704). 900 µl of the lysate 113 is transported from the lysis chamber 101 via the membrane chamber 108 to the waste liquid chamber 107 for 1 minute at an applied pressure of -60 kPa (step 705). The transported amount (900 µl) of the lysate 113 is controlled by the computer 115 based on the liquid level detection sensor 314 and the application time (1 minute) of a predetermined pressure (60 kPa). (iv) Process IV

[0068] In process IV, valves 304 and 305 are closed and valves 302, 303, and 315 are opened (step 706) before the air following the lysate 113 has passed beyond the branching point (connection point 316) of flow paths 306 and 308 and flow path 307. 500 µl of the wash buffer 111 are then transported from the wash buffer chamber 105 via the membrane chamber 108 to the waste liquid chamber 107 for 3 minutes at an applied pressure of 60 kPa (step 707). The transported quantity (500 µl) of the wash buffer 111 is controlled by the computer 115 based on the liquid level detection sensor 314 and the application time (3 minutes) of the predetermined pressure (60 kPa).

[0069] By performing the operations shown in processes I to IV, continuous transport can be achieved without trapping air between the lysate 113 and the wash buffer 111. <vergleichsbeispiel>

[0070] Fig. Figure 8 is a diagram explaining a cleaning process according to a comparative example. Fig. 9 is a flowchart of the cleaning process according to Fig. 8. (i) Process I

[0071] Process I shows the state during or immediately after lysis in sample lysis procedure 202 (step 901). At this time, valves 302, 303, 304 and 305 are closed. (ii) Process II

[0072] In process II, valves 304 and 305 are opened (step 902). The lysate 113 is transported from the lysis chamber 101 via the membrane chamber 108 to the waste liquid chamber 107 (step 903). (iii) Process III

[0073] In process III, valves 305 and 304 are closed when the transport of lysate 113 is complete (step 904). (iv) Process IV

[0074] In process IV, valves 302, 303, and 315 are opened (step 904). Wash buffer 111 is transported from wash buffer chamber 105 via membrane chamber 108 to waste liquid chamber 107 (step 905). (v) Process V

[0075] Process V shows the state after all or a predetermined portion of the wash buffer 111 has been transported via the membrane chamber 108 to the waste liquid chamber 107.

[0076] In the cleaning process according to the comparative example, the air present in the flow paths 307 and 308 enters between the lysate 113 and the wash buffer 111. Therefore, in order for the wash buffer 111 to reach the membrane, the Laplace pressure generated between the lysate 113 and the cleaning membrane 102 must be exceeded. <Technische Wirkungen>

[0077] To confirm the technical effect of the lysate and wash buffer transport process according to the first example, a comparative experiment was conducted on the pressure required for solution transport using a simple flow path system in a case where air entered the flow path and in a case where no air entered the flow path. Note that the technical effects are discussed here based on the experimental results according to the first example. The technical effects mentioned here are similar to those of the second to fourth examples and the derived examples described later. (i) Experimental conditions

[0078] In the flow path system, a lysate prepared according to a Qiagen Qiaamp DNA (QIK) assay protocol and a QIK wash buffer (AW1) were used. The lysate and the wash buffer were each placed in a 1.5 ml tube, pressurized by a syringe pump, and transported to a polycarbonate membrane chamber. The volume of solution that the membrane chamber could hold was approximately 20 µl. The membrane chamber was divided centrally by the membrane, with the volumes in front of and behind each division being approximately 10 µl. The liquid contact area of ​​the purification membrane was set to 3.1 mm². 2 The lysate and wash buffer that had passed through the membrane chamber were recovered via a waste liquid tube installed behind the membrane chamber. The extrusion speed of the syringe pump was set to 4 ml / min.

[0079] During transport (Procedure A) according to the procedure (Comparison Example), which differs from the first example, 100 µl of the lysate, 100 µl of air, and 500 µl of the wash buffer (QIK Wash Buffer AW1) were sequentially transported to the membrane chamber, and the change in pressure over time was measured. To reproduce the transport procedure according to the first example, 100 µl of the lysate and 500 µl of the wash buffer without trapped air were continuously transported in Procedure B. A Whatman GF / F glass fiber membrane was used. (ii) Experimental results

[0080] Fig. Figure 10 is a schematic diagram showing the position of the solution during transport, and a diagram showing the pressure change measured according to the experiment described above. Fig. 10. A broken line indicates the result of procedure A (comparison example) and a solid line indicates the result of procedure B (first example).

[0081] Because the extrusion speed of the syringe pump was greater than the movement speed of the solution, the pressure increased over time to 20 kPa, just before the lysate exited the membrane, while the lysate passed through the membrane ( Fig. 10 (solid line / dashed line), step (a) → (b)).

[0082] In procedure A, the liquid level stopped moving once the lysate had passed through the membrane and air had entered. Furthermore, as the syringe pump was pressed further, air began to slowly cross the membrane at 80 kPa and above. When 180 kPa was reached, the wash buffer reached the membrane chamber ( Fig. 10 (dashed line), step (c) → (d)). It was confirmed that the wash buffer crossed the membrane, and the pressure rapidly decreased to about 30 kPa after the wash buffer was completely removed ( Fig. 10 (dashed line), step (d)).

[0083] In procedure B, the solution was not stopped when the lysate passed through the membrane and the wash buffer penetrated it ( Fig. 10 (solid line), step (b) → (c)). The pressure did not exceed 40 kPa until the wash buffer was completely removed. The required pressure for transporting the single wash buffer was confirmed separately, and the transport was completed within 40 kPa (data not shown). (iii) Discussion of the experimental results

[0084] As described above, the experimental results of Procedure A (comparative example) and Procedure B (first example) confirmed that the required pressure could be reduced to 1 / 3 by using the transport method according to the first example (the same applies to other examples described later).

[0085] The reason high pressure is not required when air enters after removal of the wash buffer is that the evaporation rate of the wash buffer is significantly higher than that of the lysate. The wash buffer (QIK, AW1) consists of 50% or more ethanol by volume.

[0086] In this example, the lysate and wash buffer were transported continuously to reduce the maximum pressure required for transporting the two liquids. The effect of this example is not limited to the kit described above. The composition of the lysate or the wash buffer may vary. If the second liquid (the wash buffer) can be completely removed by Sample Detection Procedure 205, the following requirements are specified as a combination of the first liquid (the lysate) and the second liquid, thus making it easier to achieve the pressure-reducing effect. (a) The second liquid has a higher evaporation rate than the first liquid, (b) the second liquid has a lower surface tension than the first liquid, (c) the second liquid has a higher contact angle with the membrane than the first liquid, and (d) the viscosity of the second liquid is less than that of the first liquid.

[0087] If the main component of the first liquid is water, examples of a second liquid satisfying one or more of the conditions mentioned above include a solution containing 10% or more of an alcohol having four or fewer carbon atoms. A more preferred example is a solution containing 30% or more of an alcohol having three or fewer carbon atoms. A still more preferred example is a solution containing 40% or more ethanol. Additionally, a suitable second liquid may be selected according to the composition of the first liquid, or the first liquid may be selected according to the composition of the second liquid.

[0088] According to the first example (experimental result) of the present disclosure, the required pressure can be reduced. By reducing the required pressure, a high-density purification membrane can be installed in a structurally weak flow path chip. Here, examples of the structurally weak chip include a valve and a chip bonded to each other, and a chip using a soft material. Fabricating a structurally weak chip has advantages such as reducing the cost and providing high functionality of the chip. In addition, a chemical reaction that is hindered by applying high pressure can proceed efficiently by reducing the required pressure. Furthermore, the measurement of the biomolecule can be facilitated by applying high pressure.

[0089] In addition to reducing pressure, the time required for solution transport can be shortened by using the technology of the present disclosure. By using the technology of the present disclosure, the membrane can be prevented from drying out after the lysate has passed through and before washing, thus achieving benefits such as increasing biomolecule yield and washing efficiency. (iv) Membranes for DNA extraction

[0090] In the Biomolecule Analyzer 100, highly efficient DNA recovery can be achieved by installing a high-density membrane. The "particle retention size" is an index of pore size. This particle retention size means that particles with a diameter equal to or greater than the value specified by the particle retention size are retained in the membrane. In other words, the smaller the particle retention size value, the finer the membrane pores. The membrane used as a purification membrane can have a particle retention size of 100 µm or higher. For example, it is desirable to use a membrane with finer pores than the "Fusion 5" membrane distributed by Whatman. Examples of membranes capable of recovering DNA with higher efficiency include one distributed by Whatman called "GF / D." The particle retention size of this membrane is 2.7 µm.Examples of membranes capable of extracting DNA with higher efficiency include one marketed by Whatman and designated "GF / F". The particle retention size of this membrane is 0.7 µm.

[0091] Highly efficient DNA extraction can be achieved by layering multiple membranes. In the case of GF / F, for example, DNA can be extracted more efficiently by layering two or more membranes than by using a single membrane. The membrane thickness is ideally 0.001 mm or greater. In the case of GF / F, the membrane thickness is 0.6 mm. (v) Membrane chamber 108

[0092] The volume of membrane chamber 108 is 30% of the volume of lysis chamber 101 or wash buffer chamber 105, or less. The cross-sectional area in the direction perpendicular to the flow direction of the flow path through which the two liquids usually pass is 50% of the maximum cross-sectional area of ​​the chamber or less. The volume of the flow path through which the two liquids usually pass is 50% or less with respect to each of the two liquids. Therefore, the lysate and the wash buffer are mixed by no more than 50% upstream of the waste liquid chamber. Therefore, the binding efficacy of DNA is not substantially reduced even when the solution is transported using the technology of this disclosure.

[0093] The size of the membrane chamber 108 is 0.1 µl to 1 ml. Because the membrane chamber 108 is smaller, the loss during elution is smaller. Therefore, a more suitable size of the membrane chamber 108 is desirably 0.1 µl to 100 µl, and more preferably 0.1 µl to 30 µl. (vi) Flow path

[0094] The volume of the flow path 307 is 0.1 µL to 1 L (if a long flow path is used, the flow path volume increases). If the volume of the flow path 307 is larger than 1 L, solution transport takes time. Conversely, if the volume is small, the width of the flow path must be reduced or the distance between the chambers must be narrowed, and the degree of freedom in design decreases. If a chamber is located outside the chip, the volume is desirably large. If the flow path is too thin, there is a concern that an unexpected process such as flow path clogging may occur. A more suitable flow path volume is 1 µL to 100 ml, and in another example, 30 µL to 100 ml.

[0095] The volume of the flow path 307 may be equal to or greater than the volume of a space 108_1 above the cleaning membrane 102 in the membrane chamber 108.

[0096] If drying occurs after cleaning, it is desirable to ensure a certain thickness of the flow path. In this case, the thickness of flow paths 302 and 303 should ideally be 0.03 mm. 2 . <prozesswechselsteuerung>(i) Control using a liquid level detection sensor

[0097] A liquid level detection sensor can be used to switch between processes I, II, III, IV and V in Fig. 6. For example, liquid level detection sensors 313 and 314 are used when switching from process III to process IV. Sensors that detect electrically, optically, by sound waves, or dynamically can be used as liquid level detection sensors 313 and 314.

[0098] If the liquid level detection sensor 313 is used, when switching from process III to process IV, a signal obtained when the liquid level in the waste liquid chamber 107 is detected to exceed the range of the liquid level detection sensor 313 may be used. Note that the liquid level detection sensor 313 must be installed at a position where the liquid level in the waste liquid chamber 107 can be detected at the time when the residual ratio of the lysate 113 in the lysis chamber 101 becomes less than 10%.

[0099] If the liquid level detection sensor 314 is used, a signal can be used when switching from process procedure II to process procedure III. This signal is obtained when it is detected that the liquid level in the lysis chamber falls below the range of the liquid level detection sensor 314. It should be noted that the liquid level detection sensor 314 must be installed in a position where the liquid level can be detected at the time when the residual proportion of lysate 113 in the lysis chamber 101 becomes less than 10%. (ii) Control by pressure, time and the like

[0100] A pre-defined pressure and time can be used to switch between processes I, II, III and IV in Fig. 6. The pressure and time can be adjusted by the operator. A viscosity measuring mechanism may be provided in the lysis chamber 101 or the like.

[0101] A flow rate sensor or a pressure sensor can be installed at a specific location to establish a process changeover time. [Second example]

[0102] A second example will be given with reference to the Fig. 7, Fig. 11 and Fig. 12 described. <Konfigurationsbeispiel des Reinigungssystems 301, das einen Hauptteil des Strömungsweg-Chips 114 aufweist>

[0103] Fig. Figure 11 is a diagram showing a configuration example of a cleaning system 400, which has a main part of the flow path chip 114 according to the second example, i.e., a main part of the flow path chip 114 at the completion or execution of the sample lysis procedure 202.

[0104] The purification system 400 includes a lysis chamber 101, a wash buffer chamber 105, a waste liquid chamber 107, a membrane chamber 108, valves 401, 402, 403, 404, 405, and 406, flow paths 407, 408, 409, 410, 411, 412, and 413 connecting the chambers, and an orifice 109. The number of orifices 109 may not be 1, and one orifice may be installed for each function. Alternatively, for example, dedicated orifices may be provided for flow paths 413 and 412, respectively. The lysis chamber 101 receives the lysate 113. The wash buffer chamber 105 receives the wash buffer 111.

[0105] As in Fig. As shown in Figure 11, the flow path 407 originating from the outlet opening of the lysis chamber 101 leads to the inlet opening of the membrane chamber 108. The flow path 408 originating from the outlet opening of the membrane chamber 108 and the flow path 409 originating from the outlet opening of the wash buffer chamber 105 are joined at a connection point (merging point) 414, so that the flow path 410 is formed, which leads to an inlet opening on a lower section (bottom surface, which is not necessarily the bottom surface) of the waste liquid chamber 107. The flow path originating from opening 109 splits at a junction 415 into flow path 412 and flow path 413. Flow path 413 leads to an inlet opening of the wash buffer chamber 105. Flow path 412 leads to an upper section (of the upper surface, although this is not necessarily the top surface) of the waste liquid chamber 107.The flow path 411 also leads to an upper section (the upper surface, although it is not necessarily an upper surface) of the waste liquid chamber 107.

[0106] As in Fig. As shown in Figure 11, valve 401 is located in flow path 407. Valve 402 is located in flow path 409. Valve 403 is located in flow path 410. Valve 404 is located in flow path 411. Valve 405 is located in flow path 412. Valve 406 is located in flow path 413. <reinigungsprozess>

[0107] Fig. Figure 12 is a diagram explaining cleaning processes I to V according to the second example. The flowchart of the cleaning processes is similar to the one from Fig. 7. (i) Process I

[0108] Process I shows the state during lysis or immediately after completion of lysis in the sample lysing procedure 202. At this time, valves 401, 402, 403, 404, 405 and 406 are closed. (ii) Process II

[0109] In Process II, valves 402, 406, 403, and 404 are opened. The wash buffer 111 is drawn from the wash buffer chamber 105 at a pressure of no more than 60 kPa for a maximum of 30 seconds. During this time, the air in flow path 409 moves to flow path 408 or 410, and flow path 409 is filled with the wash buffer 111. As described above, expelling the air in flow path 409 in Process II is important to achieve continuous transport without trapping air between the lysate 113 and the wash buffer 111. (iii) Process III

[0110] In process III, valves 402 and 406 are closed, and valves 401, 403, and 405 are opened. 900 µl of lysate 113 are transported for 2 minutes at a pressure of -60 kPa from lysis chamber 101 via membrane chamber 108 to waste liquid chamber 107. (iv) Process IV

[0111] In process IV, valves 403 and 405 are closed and valves 402 and 406 are opened as soon as all the lysate 113 has been transported to membrane chamber 108. 500 µl of wash buffer 111 are transported from wash buffer chamber 105 via membrane chamber 108 to lysis chamber 101 for 3 minutes at a pressure of 60 kPa. (v) Process V

[0112] Process V shows the state in the middle of the transport of the washing buffer 111 from the washing buffer chamber 105 via the membrane chamber 108 to the lysis chamber 101. (vi) Process VI

[0113] Process VI shows the state after the entire wash buffer 111 or a predetermined portion thereof has been transported from the wash buffer chamber 105 via the membrane chamber 108 to the lysis chamber 101.

[0114] By using the working procedure described above, the lysate 113 and the wash buffer 111 can be transported continuously without trapping air between the lysate 113 and the wash buffer 111.

[0115] Naturally, pressure and time are also a factor in Process III. Fig. 12. The numerical values ​​mentioned above are not limited to the above-mentioned values. A combination of time and pressure must be present at which at least 90% of the lysate 113 can be transported. The time and pressure required to transport almost all of the lysate 113 from the lysis chamber 101 to the purification membrane 102 can be determined separately in an experiment and specified in an implementation program. [Third example]

[0116] A third example will be given with reference to the Fig. 13, Fig. 14 and Fig. 15 described. <Konfigurationsbeispiel des Reinigungssystems 301, das einen Hauptteil des Strömungsweg-Chips 114 aufweist>

[0117] Fig. Figure 13 is a diagram showing a configuration example of a cleaning system 500, which has a main part of the flow path chip 114 according to the third example, i.e., a main part of the flow path chip 114 at the completion or execution of the sample lysis procedure 202.

[0118] The cleaning system 500 comprises a lysis chamber 101, a wash buffer chamber 105, a waste liquid chamber 107, a membrane chamber 108, valves 501, 502, 503, 504, and 505, flow paths 506, 507, 508, 509, 510, and 511 connecting the chambers, and an opening 109. The number of openings 109 cannot be one, and one opening can be installed for each function. For example, dedicated openings can be provided in the flow paths 510 and 511, respectively. The lysis chamber 101 receives the lysate 113. The wash buffer chamber 105 receives the wash buffer 111.

[0119] As in Fig. As shown in Figure 13, the flow path 506, originating from the outlet of the lysis chamber 101, leads to one of the two inlet openings (of the upper section or upper surface, which is not necessarily an upper surface) of the membrane chamber 108. The flow path 508, originating from the outlet of the membrane chamber 108, leads to an inlet opening in the lower section (bottom surface, which is not necessarily a bottom surface) of the waste liquid chamber 107. The flow path 507, originating from the outlet of the wash buffer chamber 105, leads to the other inlet opening of the membrane chamber 108. The flow path 511, originating from the opening 109, leads to an inlet opening of the wash buffer chamber 105. The other flow path 510, originating from the opening 109, leads to an upper section (of the upper surface, which is not necessarily an upper surface). necessarily an upper surface) of the waste liquid chamber 107.The flow path 509 also leads to an upper section (the upper surface, although it is not necessarily an upper surface) of the waste liquid chamber 107.

[0120] As in Fig. As shown in Figure 13, valve 501 is located in flow path 506. Valve 502 is located in flow path 507. Valve 503 is located in flow path 509. Valve 504 is located in flow path 510. Valve 505 is located in flow path 511. <reinigungsprozess>

[0121] Fig. Figure 14 is a diagram explaining the cleaning processes I to VI according to the third example. Fig. 15 is a flowchart that corresponds to the one in Fig. This corresponds to the cleaning process shown in section 14. (i) Process I

[0122] Process I shows the state during or immediately after lysis in sample lysis procedure 202 (step 1501). At this time, valves 501, 502, 503, 504, and 505 are closed. (ii) Process II

[0123] In process II, valves 501 and 504 are opened (step 1502). 900 µl of lysate 113 are transported for 2 minutes at a pressure of -60 kPa from lysis chamber 101 via membrane chamber 108 to waste liquid chamber 107 (step 1503). (iii) Processes III and IV

[0124] In process III, valve 504 is closed and valves 502 and 505 are opened as soon as all the lysate 113 has been transported to membrane chamber 108 (step 1504). The wash buffer 111 is transported from the wash buffer chamber 105 via the membrane chamber 108 to the flow path 506 for 30 seconds at an applied pressure of 30 kPa (step 1505). During this time, the air located in chamber 108_1 on the inlet side of membrane chamber 108 moves to the flow path 506, and chamber 108_1 of membrane chamber 108 is filled with the wash buffer 111. A chamber 108_2 on the outlet side of membrane chamber 108 is filled with the lysate 113. As described above, in process IV it is important to expel the air in the membrane chamber 108 to the flow path 506 in order to achieve continuous transport without trapping the air between the lysate 113 and the wash buffer 111. (iv) Process V

[0125] When the wash buffer begins to enter the flow path 506, valve 501 is closed and valve 503 is opened (step 1506). The wash buffer 111 is transported from the wash buffer chamber 105 via the membrane chamber 108 to the waste liquid chamber 107 (step 1507). (v) Process VI

[0126] Process VI shows the state after the entire wash buffer 111 or a predetermined proportion of it has been transported from the wash buffer chamber 105 via the membrane chamber 108 to the waste liquid chamber 107.

[0127] By using the working procedure described above, the lysate 113 and the wash buffer 111 can be transported continuously without trapping air between the lysate 113 and the wash buffer 111. [Fourth example]

[0128] A fourth example is given with reference to the Fig. 15, Fig. 16 and Fig. 17 described. <Konfigurationsbeispiel des Reinigungssystems 600, das einen Hauptteil des Strömungsweg-Chips 114 aufweist>

[0129] Fig. Figure 16 is a diagram showing a configuration example of a cleaning system 600, which has a main part of the flow path chip 114 according to the fourth example, i.e., a main part of the flow path chip 114 at the completion or execution of the sample lysis procedure 202.

[0130] The cleaning system 600 comprises a lysis chamber 101, a wash buffer chamber 105, a waste liquid chamber 107, a membrane chamber 108, valves 601, 602, 603, 604, 605, and 606, flow paths 607, 608, 609, 610, 611, 612, and 613 connecting the chambers, and an opening 109. The number of openings 109 cannot be one, and one opening can be provided for each function. For example, dedicated openings can be provided for flow paths 612 and 613, respectively. The lysis chamber 101 contains the lysate 113, and the wash buffer chamber 105 contains the wash buffer 111.

[0131] As in Fig. As shown in Figure 16, the flow path 606, originating from the outlet of the lysis chamber 101, and the flow path 608, originating from the outlet of the wash buffer chamber 105, are joined at a junction point 614, forming the flow path 308. The flow path 609 leads to an inlet opening on an upper section (top surface, which is not necessarily a top surface) of the membrane chamber 108. The flow path 610, originating from an outlet opening provided in the lower section (bottom surface, which is not necessarily a bottom surface) of the membrane chamber 108, leads to an inlet opening provided in a lower section (bottom surface, which is not necessarily a bottom surface) of the waste liquid chamber 107.Flow path 611, originating from an outlet opening provided in an upper section (the upper surface, which is not necessarily an upper surface) of the membrane chamber 108, leads to an upper section (a side surface, which is not necessarily a side surface and could also be an upper surface) of the waste liquid chamber 107. Flow path 613, originating from opening 109, leads to an inlet opening of the wash buffer chamber 105. The other flow path 612, originating from opening 109, leads to an upper section (the upper surface, which is not necessarily an upper surface) of the waste liquid chamber 107. Flow path 615 also leads to an upper section (the upper surface, which is not necessarily an upper surface) of the waste liquid chamber 107.

[0132] As in Fig. As shown in Figure 16, valve 601 is located in flow path 607. Valve 602 is located in flow path 608. Valve 603 is located in flow path 615. Valve 604 is located in flow path 611. Valve 605 is located in flow path 612. Valve 606 is located in flow path 613. <reinigungsprozess>

[0133] Fig. Figure 17 is a diagram explaining cleaning processes I to V according to the fourth example. The flowchart of the cleaning process is similar to the one from Fig. 15. (i) Process I

[0134] Process I shows the state during lysis or immediately after completion of lysis in the sample lysing procedure 202. At this time, valves 601, 602, 603, 604, 605 and 606 are closed. (ii) Process II

[0135] In process II, valves 601 and 605 are opened. 900 µl of the lysate 113 are transported from the lysis chamber 101 via the membrane chamber 108 to the waste liquid chamber 107 for 2 minutes at an applied pressure of -60 kPa. (iii) Process III

[0136] In process III, valves 601 and 605 are closed and valves 602, 606, 604, and 603 are opened as soon as all the lysate 113 has been transported to the membrane chamber 108. The wash buffer 111 is transported from the wash buffer chamber 105 via the membrane chamber 108 to the flow path 611 for 30 seconds at an applied pressure of 30 kPa. (iv) Process IV and Process V

[0137] In process IV, valve 604 closes as soon as the wash buffer 111 enters the flow path 611. The wash buffer 111 is transported from the wash buffer chamber 105 via the membrane chamber 108 to the waste liquid chamber 107. At this time, the air located in chamber 108_1 on the inlet side of the membrane chamber 108 moves via the flow path 611 to the waste liquid chamber 107, and chamber 108_1 of the membrane chamber 108 is filled with the wash buffer 111. A chamber 108_2 on the outlet side of the membrane chamber 108 is filled with the lysate 113. As described above, in process IV it is important to expel the air in the membrane chamber 108 via the flow path 611 to the waste liquid chamber 107 in order to achieve continuous transport without trapping the air between the lysate 113 and the wash buffer 111. (v) Process VI

[0138] Process VI shows the state after the entire wash buffer 111 or a predetermined proportion of it has been transported from the wash buffer chamber 105 via the membrane chamber 108 to the waste liquid chamber 107.

[0139] By using the above-described working procedure, the lysate 113 and the wash buffer 111 can be transported continuously without trapping air between the lysate 113 and the wash buffer 111. [Fifth example]

[0140] A fifth example is a derived form of the first example. A cleaning system 301' according to the fifth example has a configuration in which a valve 701 is connected to the cleaning system 301 (see Fig. 6) is added according to the first example. <reinigungsprozess>

[0141] Fig. Figure 18 is a diagram explaining the cleaning processes I to V according to the fifth example. (i) Process I

[0142] Process I shows the state during or immediately after completion of lysis in sample lysis procedure 202. At this time, valves 302, 303, 304, 305 and 701 are closed. (ii) Process II

[0143] In process II, valves 305, 304, and 701 are opened. 900 µl of lysate 113 are transported for 1 minute at a pressure of -60 kPa from lysis chamber 101 via membrane chamber 108 to waste liquid chamber 107. (iii) Process III

[0144] In process III, valves 304 and 701 are closed before the air 702 following the transported lysate 113 has flowed beyond the branching point (connection point 316 of the flow paths) of flow paths 306 and 308 and flow path 307, and valves 302 and 303 are opened. The wash buffer 111 is transported from the wash buffer chamber 105 to the lysis chamber 101 via flow path 306 for 30 seconds at an applied pressure of 30 kPa. During this time, the air in flow path 307 moves to flow path 308 or 306, and flow path 307 is filled with the wash buffer 111. The expulsion of air in the flow path 307 in process III is important to achieve continuous transport without trapping air between the lysate 113 and the wash buffer 111. (iv) Process IV

[0145] In process IV, valves 315 and 701 are opened. 500 µl of the wash buffer 111 are transported for 3 minutes at 60 kPa from the wash buffer chamber 105 via the membrane chamber 108 to the waste liquid chamber 107. (v) Process V

[0146] Process V shows the state after the entire wash buffer 111 or a predetermined portion thereof has been transported from the wash buffer chamber 105 via the membrane chamber 108 to the waste liquid chamber 107.

[0147] By using the above-described working procedure, the lysate 113 and the wash buffer 111 can be transported continuously without trapping air between the lysate 113 and the wash buffer 111. [Sixth example]

[0148] A sixth example is a derivative of the second example. A cleaning system 400' according to the sixth example has a configuration in which a valve 801 is connected to the cleaning system 400 (see Fig. 11) as shown in the second example. <reinigungsprozess>

[0149] Fig. Figure 19 is a diagram explaining the cleaning processes I to V according to the sixth example. (i) Process I

[0150] Process I shows the state during or immediately after completion of lysis in sample lysis procedure 202. At this time, valves 401, 402, 403, 404, 405, 406 and 801 are closed. (ii) Process II

[0151] In process II, valves 401, 403, 405, and 801 are opened. 900 µl of lysate 113 are transported for 2 minutes at a pressure of -60 kPa from lysis chamber 101 via membrane chamber 108 to waste liquid chamber 107. (iii) Process III

[0152] In process III, valves 405 and 801 are closed and valves 402, 406, and 404 are opened as soon as all the lysate 113 has been transported to membrane chamber 108. The wash buffer 111 is transferred from wash buffer chamber 105 to waste liquid chamber 107 at a pressure of 30 kPa or less over a period of 30 seconds or less. During this time, the air in flow path 409 moves to flow path 410 or to waste liquid chamber 107. Alternatively, the air in flow path 409 can move to flow path 408 or 410, and flow path 409 is filled with the wash buffer 111. As described above, in process III, it is important to expel the air in the flow path 409 to achieve continuous transport without trapping the air between the lysate 113 and the wash buffer 111. (iv) Process IV

[0153] In process IV, valve 403 is closed and valve 801 is opened. 500 µl of wash buffer 111 are transported from wash buffer chamber 105 via membrane chamber 108 to lysis chamber 101 for 3 minutes at 60 kPa. (v) Process V

[0154] Process V shows the state after all or a predetermined portion of the wash buffer 111 has been transported from the wash buffer chamber 105 via the membrane chamber 108 to the lysis chamber 101.

[0155] By using the working procedure described above, the lysate 113 and the wash buffer 111 can be transported continuously without trapping air between the lysate 113 and the wash buffer 111. (3) Supplementary description of examples

[0156] (i) In the first example, for continuous transport of the washing buffer and the lysate, a switch from process III to process IV must be made at the correct time, as shown in Fig. Figure 6 illustrates this. If the transport time of lysate 113 is too long, air enters the flow path 308. If the transport time is too short, a large amount of lysate 113 remains in the lysis chamber 101, thus reducing the yield. Therefore, the transport must be carried out with high reproducibility, or the transport time must be defined by the liquid level detection sensor.

[0157] However, in cases where the sample type varies or the liquid viscosity is high, the reproducibility of lysate 113 transport is low. Using the liquid level detection sensor increases costs due to the increased number of device components. There is also a risk that the liquid level detection sensor will malfunction.

[0158] On the other hand, when switching from Process III to Process IV in the second example, as in Fig. 11 shown, during the switch from process II to process III in the third example, as in Fig. 14 shown, and when switching from process II to process III in the fourth example, as in Fig. As shown in Figure 17, the wash buffer 111 and the lysate 113 are transported continuously even if the time required for the transport of the lysate 113 is exceeded. Therefore, in these examples, even if no liquid level detection sensor is used in the first example, a configuration can be achieved in which the liquid level detection sensor is unnecessary while maintaining the yield.

[0159] (ii) The lysate 113 includes a sample-derived polymer, a particle, a precipitate derived from a mixture with a lysis buffer, and the like. These are captured by the purification membrane during transport of the lysate 113, which may lead to a reduction in the efficiency of transporting the wash buffer 111 and the eluate at the subsequent stage or a reduction in the efficiency of transporting the fluid (air, nitrogen, and other gases) during drying.

[0160] Therefore, in the third example, the washing buffer 111 is caused to flow from the side opposite the lysate 113. This also ensures that during transport, pressure is exerted against any object trapped at the membrane, thus achieving smooth transport.

[0161] (iii) In the third example, the fourth example, the fifth example (a derivative of the first example), and the sixth example (a derivative of the second example), 4 it is not necessary to divide the transport of the washing buffer 111 into multiple times. Therefore, the washing buffer chamber 105 does not need to have a structure corresponding to multiple transport times. For example, a reagent transport package using an actuator as shown in US 2006 / 134773 A1 may be used as the reagent chamber. However, in the fifth example (a derivative of the first example) and the sixth example (a derivative of the second example), unless the valve 701 or the valve 801 is provided, there is a possibility that the fluid (air, nitrogen, and other gases) moves to the membrane chamber 108 side.Therefore, although transport is possible without it, valve 701 or 801 can be provided to achieve even more stable solution transport. The flow path according to the third example is usually longer than that in the first example. The flow path according to the fourth example requires an additional flow path beyond that of the first example. (4) Summary of examples

[0162] In each example, in a flow path with a purification membrane, two types of solutions can be continuously transported to a location where the membrane is installed without installing a dedicated bubble removal configuration in a flow path chip, and the pressure required for solution transport can be reduced. This allows the number of gas-liquid interfaces with a high Laplace pressure to be reduced or zeroed. Furthermore, the volume of fluid (air, nitrogen, and other gases) that needs to be transported in a wet state of the membrane can be reduced or zeroed.

[0163] (i) According to the first example (see Fig. 5 and Fig. 6) The purification system (biomolecule analyzer) 301 has a flow path in which the membrane chamber 108 is located between the lysis chamber (first chamber) 101 and the wash buffer chamber (second chamber) 105 and the waste liquid chamber 107. The computer 115 controls the transport of the wash buffer (second liquid) 111 until the wash buffer has flowed past at least the junction point 316 between the flow paths 306 to 309 (first flow path) leading from the lysis chamber 101 to the waste liquid chamber 107 and the flow path 307 (second flow path) leading from the wash buffer chamber (second chamber) 105, and expels the fluid (air, nitrogen, and other gases) from the flow path 307 (second flow path). Next, the computer 115 controls the transport of the lysate (first liquid) 113 from the lysis chamber (first chamber) 101 to the waste liquid chamber 107 via the membrane chamber 108.Then, the computer 115 controls the transport of the wash buffer (second liquid) 111 in the wash buffer chamber (second chamber) 105 from the wash buffer chamber (second chamber) 105 to the waste liquid chamber 107 via the membrane chamber 108. This allows the continuous transport of the lysate 113 and the wash buffer 111 to the waste liquid chamber 107 without air mixing during the solution transport between the lysate 113 and the wash buffer 111. Because the fluid (air, nitrogen, and other gases) is not mixed, the pressure can be reduced during the solution transport (see ). Fig. 10). Therefore, even if the flow path is structurally somewhat weak, it can be used, and the manufacturing cost of the cleaning system 301 can be reduced.

[0164] (i) According to the second example (see Fig. 11 and Fig. 12), the purification system (biomolecule analyzer) 400 has the flow path in which the membrane chamber 108 is located between the lysis chamber (first chamber) 101 and the wash buffer chamber (second chamber) 105. The membrane chamber 108 and the wash buffer chamber 105 are connected to the waste liquid chamber 107. The computer 115 controls the transport of the wash buffer (second liquid) 111 until the wash buffer has passed at least beyond the junction point 414 between the flow paths 407 to 408 (first flow path) leading from the lysis chamber (first chamber) 101 to the waste liquid chamber 107 and the flow path 409 (second flow path) leading from the wash buffer chamber (second chamber) 105, and expels the fluid (air, nitrogen, and other gases) from the flow path 409 (second flow path).Next, the computer 115 controls the transport of the lysate (first liquid) 113 from the lysis chamber (first chamber) 101 to the waste liquid chamber 107 via the membrane chamber 108. Then, the computer 115 controls the transport of the wash buffer (second liquid) 111 in the wash buffer chamber (second chamber) 105 from the wash buffer chamber (second chamber) 105 to the lysis chamber (first chamber) 101 via the membrane chamber 108. This enables the transport of the lysate 113 to the waste liquid chamber 107 and then the continuous transport of the wash buffer 111 to the lysis chamber 101 without interfering with any fluid (air, nitrogen, and other gases) between the lysate 113 and the wash buffer 111 during solution transport. Because the fluid (air, nitrogen and other gases) is not mixed, the pressure can be reduced during solution transport (similar to the first example, see . Fig. 10) Therefore, even if the flow path is structurally somewhat weak, it can be used and the manufacturing costs of the cleaning system 400 can be reduced.

[0165] (iii) According to the third example (see Fig. 13 and Fig. 12), the purification system (biomolecule analyzer) 500 has a flow path in which the membrane chamber has an inlet port leading to the lysis chamber (first chamber) 101, a second inlet port leading to the wash buffer chamber (second chamber) 105 on the upstream side, and an outlet port leading to the waste liquid chamber 107. The flow path 506 (first flow path), which originates from the lysis chamber (first chamber) 101, leads to the first inlet port of the membrane chamber 108. The flow path 507 (second flow path), which originates from the wash buffer chamber (second chamber) 105, leads to the second inlet port of the membrane chamber 108. The flow path 508 (third flow path), which originates from the outlet port of the membrane chamber 108, leads to the waste liquid chamber 107. The computer 115 transports the lysate (the first liquid) 113 from the lysis chamber (first chamber) 101 via the membrane chamber 108 to the waste liquid chamber 107.Next, the computer 115 transports the washing buffer (the second liquid) from the washing buffer chamber (second chamber) 105 via the second inlet port of the membrane chamber 108 to the space 108_1 between the first and second inlet ports of the membrane chamber 108 and the cleaning membrane of the membrane chamber 108, and discharges the washing buffer (the second liquid) 111 from the first inlet port of the membrane chamber 108 to the flow path 506 (first flow path) side in a state where the space 108_1 is filled with the washing buffer (the second liquid) 111. Then the computer 115 transports the wash buffer (the second liquid) in a state in which the wash buffer (the second liquid) 111 is located on the side of the flow path 506 (of the first flow path) from the wash buffer chamber (second chamber) via the membrane chamber 108 to the waste liquid chamber 107.This allows the continuous transport of the lysate 113 and the wash buffer 111 to the waste liquid chamber 107 without mixing any fluid (air, nitrogen, and other gases) during solution transport between the lysate 113 and the wash buffer 111. Because the fluid (air, nitrogen, and other gases) are not mixed, the pressure can be reduced during solution transport (similar to the first example, see ). Fig. 10). Therefore, even if the flow path is structurally somewhat weak, it can be used and the manufacturing costs of the cleaning system 500 can be reduced.

[0166] (iv) According to the fourth example (see Fig. 16 and Fig. 17) The purification system (the biomolecule analyzer) 600 has a flow path in which a membrane chamber 108 has a purification membrane 102, an inlet opening through which a lysis chamber (first chamber) 101 and a washing buffer chamber (second chamber) 105 are connected via a flow path 609, a communication opening with a waste liquid chamber 107 and an outlet opening.The flow path of the purification system (biomolecule analyzer) 600 comprises flow paths 607 to 609 (the first flow path), which originate from the lysis chamber (first chamber) 101 and lead to the inlet opening of the membrane chamber 108, a flow path 608 (second flow path), which extends from the washing buffer chamber (second chamber) 105 to a merging point 614 on the flow paths 607 to 609 (the first flow path), a flow path 611 (communication flow path), which leads to the communication opening provided in a first space 108_1 between the inlet opening of the membrane chamber 108 and the purification membrane and the waste liquid chamber 107, and a flow path 610 (third flow path), which is provided from the outlet opening of the membrane chamber 108 and leads to the waste liquid chamber 107.The computer 115 transports the lysate (first liquid) 113 from the lysis chamber (first chamber) 101 via the membrane chamber 108 to the waste liquid chamber 107. Next, the computer 115 transports the wash buffer (second liquid) 113 through the inlet port of the membrane chamber 108 from the wash buffer chamber (second chamber) 105 to the first space 108_1 and the flow path 611 (communication flow path). Then, the computer 115 transports the wash buffer (second liquid) 111 from the wash buffer chamber (second chamber) 105 via the membrane chamber 108 to the waste liquid chamber 107 in a state where the first space 108_1 and the flow path 611 (communication flow path) are filled with the wash buffer (second liquid) 111.This enables the continuous transport of the lysate 113 and the wash buffer 111 to the waste liquid chamber 107 without mixing in any fluid (air, nitrogen, and other gases) during the solution transport between the lysate 113 and the wash buffer 111. Because the fluid (air, nitrogen, and other gases) is not mixed, the pressure can be reduced during the solution transport (similar to the first example, see Figure 1). Fig. 10). Therefore, even if the flow path is structurally somewhat weak, it can be used and the manufacturing costs of the 600 cleaning system can be reduced. List of reference symbols 100 Biomolecule Analyzer 101 Lysis Chamber 102 cleaning membrane 103 Reaction chamber 104 Lysis buffer chamber 105 Wash buffer chamber 106 Reagent chamber 107 Waste liquid chamber 108 Membrane chamber 109 Opening 114 Flow path chip 115 computers 301, 301', 400, 400', 500, 600 cleaning system 313, 314 Liquid level detection sensor QUOTES CONTAINED IN THE DESCRIPTION

[0000] This list of documents submitted by the applicant was generated automatically and is included solely for the convenience of the reader. This list is not part of the German patent or utility model application. The DPMA assumes no liability for any errors or omissions. Cited patent literature

[0000] US 10233491

[0006] US 9354199

[0006] JP 6613212

[0006] US 2006 / 134773 A1

[0161] Cited non-patent literature

[0000] PDMS membranes with tunable gas permeability for microfluidic applications - RSC Advances (RSC Publishing) DOI:10.1039 / C4RA1293B", RSC Adv., 2014, 4, 61415

[0006] Integrated Microfluidic System for Rapid Forensic DNA Analysis: Sample Collection to DNA Profile / Analytical Chemistry (acs.org)", Anal. Chem. 2010, 82, 16, 6991-6999

[0006] < / reinigungsprozess> < / reinigungsprozess> < / reinigungsprozess> < / reinigungsprozess> < / reinigungsprozess> < / prozesswechselsteuerung> < / vergleichsbeispiel> < / reinigungsprozess> < / nachweisverfahren> < / reinigungsmembran> < / waschpuffer> < / lysat> < / probentyp> < / reinigungssystem> < / kammer>

Claims

[1] A method for controlling the liquid transport in a flow path of a biomolecule analyzer by a computer, wherein the biomolecule analyzer comprises a first chamber storing a first liquid, a second chamber storing a second liquid, a membrane chamber with a cleaning membrane, and a waste liquid chamber, the method comprising: Controlling the transport of the second liquid until the second liquid has flowed at least beyond a junction point between a first flow path leading from the first chamber to the waste liquid chamber and a second flow path leading from the second chamber, and expelling a fluid different from the first and second liquids from the second flow path by the computer, Controlling the transport of the first liquid from the first chamber to the waste liquid chamber via the membrane chamber by the computer and Controlling the transport of the second fluid in the second chamber from the second chamber to the membrane chamber by the computer. [2] The method according to claim 1, wherein the membrane chamber is arranged between the first and second chambers and the waste liquid chamber and the method further comprises: Controlling the further transport of the second liquid transported to the membrane chamber to the waste liquid chamber by the computer. [3] The method according to claim 1, wherein the membrane chamber is arranged between the first and second chambers and the method further comprises: Controlling the further transport of the second liquid transported to the membrane chamber to the first chamber by the computer. [4] The method of claim 1, wherein the computer is configured to control the applied pressure and the opening and closing of a valve in response to input information about flow rates of the first and second liquids and to carry out each transport of the first and second liquids. [5] Method according to claim 4, wherein the membrane chamber is arranged between the first and second chambers and the waste liquid chamber and the computer is designed to (i) transport the second liquid by closing a first valve provided closer to the first chamber than the merging point in the first flow path, opening a second valve provided in the second flow path, and applying pressure to the liquid so that it overflows at least beyond the merging point, (ii) then transport the first liquid from the first chamber via the membrane chamber to the waste liquid chamber by closing the second valve, opening the first valve, and applying pressure to the liquid, and (iii) further transport the second liquid from the second chamber via the membrane chamber to the waste liquid chamber by closing the first valve, reopening the second valve, and applying pressure to the liquid. [6] Method according to claim 4, wherein the membrane chamber is arranged between the first and second chambers and the computer is designed to (i) transport the second liquid by closing a first valve provided between the first chamber and the membrane chamber, opening a second valve provided in the second flow path and a third valve provided between the merging point and the waste liquid chamber, and applying pressure to the liquid so that it overflows at least beyond the merging point, (ii) then transport the first liquid from the first chamber via the membrane chamber to the waste liquid chamber by closing the second valve, opening the first and third valves, and applying pressure to the liquid, and (iii) further transport the second liquid from the second chamber via the membrane chamber to the first chamber by closing the third valve, opening the first and second valves, and applying pressure to the liquid. [7] Method according to claim 4, wherein the membrane chamber is arranged between the first and second chambers and the waste liquid chamber and the computer is designed to (i) transport the first liquid by opening a first valve provided between the merging point and the first chamber and a second valve provided between the merging point and the membrane chamber, closing a third valve provided in the second flow path, and applying pressure to the liquid from the first chamber to the waste liquid chamber via the membrane chamber to fill a flow path from the third valve to an inlet opening of the waste liquid chamber with the first liquid, (ii) then transport the second liquid by closing the second valve, opening the first and third valves, and applying pressure to the liquid so that it overflows at least beyond the merging point, and (iii) further transport the second liquid by closing the first and second valves,Opening the third valve and applying pressure to transport the liquid from the second chamber through the membrane chamber to the waste liquid chamber. [8] Method according to claim 4, wherein the membrane chamber is arranged between the first and second chambers and the computer is designed to (i) transport the first liquid by opening a first valve provided between the first chamber and the membrane chamber, a second valve provided between the membrane chamber and the merging point, and a third valve provided between the merging point and the waste liquid chamber, closing a fourth valve provided in the second flow path, and applying a pressure to the liquid from the first chamber via the membrane chamber to the waste liquid chamber, (ii) then transport it so as to overflow at least beyond the merging point, (iii) then the second liquid by closing the second valve while maintaining a conditionin which a space between the cleaning membrane and a first liquid outlet opening of the membrane chamber and a flow path between the first liquid outlet opening and the second valve is filled with the first liquid, opening the third and fourth valves and applying pressure to the liquid to transport from the second chamber to the waste liquid chamber to expel a fluid different from the first and second liquids contained in the second flow path to the waste liquid chamber, and (iv) further transporting the second liquid from the second chamber to the first chamber by closing the third valve, opening the first, second, and fourth valves, and applying pressure to the liquid via the membrane chamber. [9] The method of claim 1, wherein the first flow path connected to the membrane chamber has a cross-sectional area of ​​314 mm 2 or less. [10] The method according to claim 1, wherein the purification membrane is capable of retaining particles of 0.1 µm or more. [11] The method of claim 1, wherein a component of the purification membrane is silica. [12] The method of claim 1, wherein the first liquid contains (i) a biopolymer containing a nucleic acid, a protein, a lipid or a polysaccharide, or a biomonomer containing an amino acid, a lipid, a sugar or a nucleobase, and (ii) a substance having a molecule having in its structure a derivative of the biopolymer or the biomonomer. [13] The method of claim 1, wherein the second liquid has a higher evaporation rate, a lower surface tension, or a higher contact angle with respect to a membrane than the first liquid. [14] The method of claim 1, wherein the first liquid is a lysate, the second liquid is a wash buffer, and the fluid is air. [15] The method of claim 1, wherein the computer is configured to transport the first and second fluids at a pressure lower than a pressure required for the fluid to pass through the purification membrane after the first fluid has passed through the purification membrane. [16] The method of claim 4, wherein the computer is configured to determine the times of alternating transport of the first and second liquids by a liquid level detection sensor, time, pressure, or a combination thereof. [17] The method of claim 1, wherein the volume of the second flow path is equal to or greater than the volume of a first space between the cleaning membrane and an inlet opening of the membrane chamber. [18] A method for controlling the liquid transport in a flow path of a biomolecule analyzer by a computer, wherein the biomolecule analyzer has a first chamber storing a first liquid, a second chamber storing a second liquid, a membrane chamber with a cleaning membrane, and a waste liquid chamber, the membrane chamber has a first inlet opening, a second inlet opening, and an outlet opening, a first flow path leading from the first chamber to the first inlet opening of the membrane chamber, a second flow path leading from the second chamber to the second inlet opening of the membrane chamber, and a third flow path leading from the outlet opening of the membrane chamber to the waste liquid chamber, the method comprising: Transporting the first liquid from the first chamber to the waste liquid chamber via the membrane chamber through the computer, Transporting the second liquid from the second chamber to a first space between the first and second inlet ports of the membrane chamber and the cleaning membrane via the second inlet port of the membrane chamber by the computer and discharging the second liquid from the first inlet port of the membrane chamber to the first flow path side in a state in which the first space is filled with the second liquid, and Transporting the second liquid from the second chamber via the membrane chamber to the waste liquid chamber by the computer in a state where the second liquid is on the side of the first flow path. [19] The method of claim 18, wherein the computer is configured to control the applied pressure and the opening and closing of a valve in response to input information about flow rates of the first and second liquids and to carry out each transport of the first and second liquids. [20] The method according to claim 19, wherein the computer is configured to (i) transport the first liquid from the first chamber to the waste liquid chamber via the membrane chamber by opening a first valve provided in the first flow path, closing a second valve provided in the second flow path, and applying pressure to the liquid; (ii) discharge the second liquid from the first inlet port to the first flow path side by opening the first and second valves and applying pressure to the liquid in a state where a second space between the cleaning membrane of the membrane chamber and the outlet port is filled with the first liquid, while filling the first space of the membrane chamber with the second liquid; and (iii) further discharge the second liquid by closing the first valve and applying pressure to the liquid in a state wherein which the second liquid is located on the side of the first flow path, from the second chamber via the membrane chamber to the waste liquid chamber., [21] A method for controlling the liquid transport in a flow path of a biomolecule analyzer by a computer, wherein the biomolecule analyzer comprises a first chamber storing a first liquid, a second chamber storing a second liquid, a membrane chamber with a cleaning membrane, and a waste liquid chamber; a first flow path leading from the first chamber to an inlet opening of the membrane chamber; a second flow path leading from the second chamber is connected to the first flow path at a junction point; and further, a communication flow path connecting a first space between the inlet opening of the membrane chamber and the cleaning membrane and the waste liquid chamber is provided; the method comprising: Transporting the first liquid from the first chamber via the membrane chamber to the waste liquid chamber through the computer, Transporting the second liquid from the second chamber to the first space and to the communication flow path from the inlet opening of the membrane chamber through the computer and Transporting the second liquid from the second chamber via the membrane chamber to the waste liquid chamber by the computer in a state where the first space and the communication flow path are filled with the second liquid. [22] The method of claim 21, wherein the computer is configured to control the applied pressure and the opening and closing of a valve in response to input information about flow rates of the first and second liquids and to carry out each transport of the first and second liquids. [23] The method according to claim 22, wherein the computer is configured to (i) transport the first liquid from the first chamber to the waste liquid chamber via the membrane chamber by opening a first valve provided in the first flow path, closing a second valve provided in the second flow path, and applying a liquid supply pressure; (ii) fill the first space of the membrane chamber and the communication flow path with the second liquid by closing the first valve in a state where a second space between the cleaning membrane and an outlet port of the membrane chamber is filled with the first liquid, and opening a second valve provided in the second flow path and a third valve provided in the communication flow path and applying a liquid supply pressure;and (iii) further transport the second liquid by closing the third valve and applying a liquid supply pressure from the second chamber via the diaphragm chamber to the waste liquid chamber while maintaining a state in which the second liquid is in the communication flow path. [24] Biomolecule purification system, comprising: a biomolecule analyzer with a flow path and a computer designed to control the fluid transport in the flow path, wherein the biomolecule analyzer has the following: a first chamber storing a first liquid, a second chamber storing a second liquid, a waste liquid chamber and a membrane chamber with a cleaning membrane arranged between the first and second chambers and the waste liquid chamber, wherein the computer is designed to perform the following: Controlling the transport of the second liquid until the second liquid has flowed at least beyond a junction point between a first flow path leading from the first chamber to the waste liquid chamber and a second flow path leading from the second chamber, and expelling a fluid different from the first and second liquids from the second flow path, Controlling the transport of the first liquid from the first chamber via the membrane chamber to the waste liquid chamber and Controlling the transport of the second liquid in the second chamber from the second chamber via the membrane chamber to the waste liquid chamber. [25] Biomolecule purification system, comprising: a biomolecule analyzer with a flow path and a computer designed to control the fluid transport in the flow path, wherein the biomolecule analyzer has the following: a first chamber storing a first liquid, a second chamber storing a second liquid, a membrane chamber with a cleaning membrane arranged between the first and second chambers, and a waste liquid chamber, wherein the computer is designed to perform the following: Controlling the transport of the second liquid until the second liquid has flowed at least beyond a junction point between a first flow path leading from the first chamber to the waste liquid chamber and a second flow path leading from the second chamber, and expelling a fluid different from the first and second liquids from the second flow path, Controlling the transport of the first liquid from the first chamber via the membrane chamber to the waste liquid chamber and Controlling the transport of the second fluid in the second chamber from the second chamber via the membrane chamber to the first chamber. [26] Biomolecule purification system, comprising: a biomolecule analyzer with a flow path and a computer designed to control the fluid transport in the flow path, wherein the biomolecule analyzer has the following: a first chamber storing a first liquid, a second chamber storing a second liquid, a membrane chamber with a cleaning membrane, a first inlet opening, a second inlet opening and an outlet opening, a waste liquid chamber, a first flow path extending from the first chamber and leading to the first inlet opening of the membrane chamber, a second flow path extending from the second chamber and leading to the second inlet port of the diaphragm chamber, and a third flow path leading from the outlet opening of the membrane chamber to the waste liquid chamber, wherein the computer is designed to perform the following: Transporting the first liquid from the first chamber via the membrane chamber to the waste liquid chamber, Transporting the second liquid from the second chamber to a first space between the first and second inlet openings of the membrane chamber and the cleaning membrane via the second inlet opening of the membrane chamber and discharging the second liquid from the first inlet opening of the membrane chamber to the first flow path side in a state in which the first space is filled with the second liquid, and Transporting the second liquid from the second chamber via the membrane chamber to the waste liquid chamber in a state where the second liquid is on the side of the first flow path. [27] Biomolecule purification system, comprising: a biomolecule analyzer with a flow path and a computer designed to control the fluid transport in the flow path, wherein the biomolecule analyzer has the following: a first chamber storing a first liquid, a second chamber storing a second liquid, a waste liquid chamber, a membrane chamber with a cleaning membrane, an inlet opening, a communication opening with the waste liquid chamber and an outlet opening, a first flow path extending from the first chamber and leading to the inlet opening of the membrane chamber, a second flow path originating from the second chamber and extending to a junction point with the first flow path, a communication flow path connecting the communication port and the waste liquid chamber, the communication flow path being provided in a first space between the inlet port of the membrane chamber and the cleaning membrane, and a third flow path leading from the outlet port of the membrane chamber to the waste liquid chamber. [28] Biomolecule purification system according to claim 27, wherein the computer is configured to perform the following: Transporting the first liquid from the first chamber via the membrane chamber to the waste liquid chamber, Transporting the second liquid from the second chamber to the first space and to the communication flow path via the inlet opening of the membrane chamber and Transporting the second liquid from the second chamber via the diaphragm chamber to the waste liquid chamber in a state where the first space and the communication flow path are filled with the second liquid.

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