EP1993730A1 - Procédé de réalisation d'une réaction de multiplication d'un acide nucléique - Google Patents

Procédé de réalisation d'une réaction de multiplication d'un acide nucléique

Info

Publication number
EP1993730A1
EP1993730A1 EP07711807A EP07711807A EP1993730A1 EP 1993730 A1 EP1993730 A1 EP 1993730A1 EP 07711807 A EP07711807 A EP 07711807A EP 07711807 A EP07711807 A EP 07711807A EP 1993730 A1 EP1993730 A1 EP 1993730A1
Authority
EP
European Patent Office
Prior art keywords
liquid
reaction vessel
piston
conducting system
vessel
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP07711807A
Other languages
German (de)
English (en)
Inventor
Jürgen SCHÜLEIN
Ugur ÜLKER
Jürgen KRAUSE
Dirk Kuhlmeier
Roland Barten
Herbert Argauer
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Kloiber Michael
Progen Biotechnik GmbH
Original Assignee
Kloiber Michael
Directif GmbH
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Kloiber Michael, Directif GmbH filed Critical Kloiber Michael
Publication of EP1993730A1 publication Critical patent/EP1993730A1/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L7/00—Heating or cooling apparatus; Heat insulating devices
    • B01L7/52—Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/04—Exchange or ejection of cartridges, containers or reservoirs
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/06—Fluid handling related problems
    • B01L2200/0684—Venting, avoiding backpressure, avoid gas bubbles
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/14—Process control and prevention of errors
    • B01L2200/142—Preventing evaporation
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0809—Geometry, shape and general structure rectangular shaped
    • B01L2300/0816—Cards, e.g. flat sample carriers usually with flow in two horizontal directions
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/18—Means for temperature control
    • B01L2300/1838—Means for temperature control using fluid heat transfer medium
    • B01L2300/1844—Means for temperature control using fluid heat transfer medium using fans
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0475—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure
    • B01L2400/0478—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure pistons
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip

Definitions

  • the invention relates to a method for carrying out a reaction for the amplification of a nucleic acid (nucleic acid amplification reaction, - NSV) in a liquid-conducting system and to a device suitable for carrying out the method.
  • a fluid-conducting system is understood to be an arrangement of fluid-conducting or fluidically interconnected fluid conduits and fluid chambers.
  • Such a method and apparatus are known in the art.
  • a liquid which contains a nucleic acid to be amplified and all reagents required for carrying out a polymerase chain reaction (PCR) is passed through a hose.
  • the tube is heated at different intervals in different sections, so that the liquid flowing through it passes through the temperatures required for the PCR.
  • a disadvantage of this method is that the temperature profile, which passes through the liquid, is determined by the course of the tube by predetermined temperature zones. It is not or only with great difficulty possible with such a device to change the temperature profile required for carrying out the PCR and thereby adapt it, for example, to the nucleic acid to be amplified.
  • the object of the present invention is to provide a method and a device which make it possible to carry out a NSI within a fluidic system while avoiding the abovementioned disadvantages.
  • liquid for example in the form of vapor, escapes from the device. This would change the defined concentration or amount of reagents or nucleic acid to be amplified present in the liquid.
  • a method for carrying out a reaction for amplifying a nucleic acid (nucleic acid amplification reaction, - NSV) in a liquid-conducting provided the system, wherein the nucleic acid is contained in a first liquid.
  • a cylindrical reaction vessel is provided which has an opening at a first end, which is connected in fluid-conducting manner via a first liquid line to the liquid-conducting system. Via the first liquid line, the reaction vessel can be filled with the first liquid and emptied. Between an outer space and the interior of the reaction vessel, the reaction vessel has a first piston displaceable in the reaction vessel.
  • the method comprises the following steps:
  • the reaction vessel is also connected to the liquid-conducting system in a liquid-conducting manner, when the flow of the liquid between the reaction vessel and the remaining liquid-conducting system is reversibly interrupted by a valve or another device.
  • the first liquid can partially replace the reagents required to carry out the NSV in addition to the nucleic acid to be amplified or completely included.
  • the first fluid may also contain only the nucleic acid.
  • the first liquid is any liquid containing the first liquid. Accordingly, a first liquid is understood as meaning a mixture of first liquid and other liquids or a solution of reagents in the first liquid.
  • the introduction of the first liquid into the reaction vessel according to step lit. a) can also be done after the first liquid has been moved back and forth between the reaction vessel and another vessel for better mixing or dissolution of reagents in the liquid.
  • a reaction vessel is provided by the first piston, which has a variable volume by the movable first piston.
  • the method according to the invention makes it possible to carry out a NSI in a fluidic system under precisely defined conditions with regard to temperature, concentration of the reagents used and amount of nucleic acid used. Preventing fluid loss through the plunger also helps prevent contamination of the infectious specimen. Furthermore, contamination of the first liquid by contaminants from the environment can also be avoided by the first piston. This is particularly important in carrying out an NSV which causes an exponential nucleic acid amplification since even the smallest impurities, for example due to dander falling into the reaction vessel, can falsify the result.
  • the first piston allows a pressure equalization between the interior of the reaction vessel and the first piston
  • the fluidic system is preferably a microfluidic system.
  • a microfluidic system is understood to mean a fluid-conducting system in which the fluid conduits have a diameter or a diagonal in a predominant part of the length of the fluid conduits, which is shorter than 2 mm, in particular shorter than 1 mm, preferably shorter than 0.5 mm , is.
  • the first piston is located at the first end of the cylindrical reaction vessel before the first liquid is introduced.
  • the volume of the interior of the reaction vessel is minimized, so that after the introduction of the first liquid no or almost no gas such.
  • B. air located in the reaction vessel.
  • a gas in the reaction vessel is compressible and thereby makes it more difficult to discharge a precisely defined volume from the reaction vessel by pressing the first piston.
  • the first piston is displaced by the introduction of the first liquid.
  • the NSV is preferably a polymerase chain reaction (PCR) or another reaction in which the temperature of the first liquid is changed. If the first liquid changes its volume or forms gas by a temperature change, either by outgassing air dissolved therein or by vaporization, the first piston can be moved by the first liquid, thereby changing the volume of the interior of the reaction vessel.
  • PCR polymerase chain reaction
  • the first piston is only exposed during the entire process by the first liquid and / or by gas formed from the first liquid and / or by pressing the first piston from outside the reaction vessel. vessel is moved.
  • a particularly simple design actuator for automated implementation of the method can be used, because no constructive measures must be taken to move the first piston by train.
  • the first piston lit. a) and / or lit. b) is moved to a predetermined stop.
  • the predetermined stop makes it possible to fill a defined volume in the reaction vessel. Given a concentration of the nucleic acid to be amplified in the first liquid, a precisely defined amount of nucleic acid to be amplified can be filled into the reaction vessel. By the stop can be ensured that enters the reaction vessel a defined volume, without the amount of liquid which is forced through the opening of the reaction vessel, must be regulated on the part of the remaining liquid-conducting system.
  • the stop between the steps lit. a) and lit. b) is displaced or removed in the opposite direction to the opening.
  • the attack can be z. B. be formed by a displaceable or removable rod.
  • a respective desired volume of liquid can be adjusted by the stop when filling the reaction vessel. After filling, the stop may be removed or withdrawn to carry out the reaction, thereby allowing pressure equalization.
  • the counterforce can be generated by a force acting on the first piston by an expansion of the first liquid against gravity to moving weight or by a frictional resistance of the first piston relative to the reaction vessel.
  • the weight can be z. B. act on the piston by the piston itself has this weight. Due to the frictional resistance of the piston moves only when a certain minimum pressure is reached and it comes to a stop before complete pressure in the reaction vessel is reached.
  • the reaction vessel is preferably arranged in such a way that bubbles forming in the first liquid can rise to the first piston.
  • This can be achieved, in particular, by arranging the longitudinal axis of the reaction vessel essentially vertically.
  • the bubbles enter the liquid-conducting system during the subsequent emptying of the reaction vessel.
  • This can be done in particular by the fact that the first piston for pushing out the first liquid in the direction of the first end but not pressed to the first end. It can be a defined
  • the method is the ratio of height to inner diameter greater than 2, preferably greater than 4, in particular greater than 6.
  • the cylindrical reaction vessel can in particular also run slightly conically towards the first end.
  • a first piston made of an elastic material this is achieved in that the first piston can easily be displaced by the introduction of the first fluid and a lower pressure is required for the introduction.
  • the wall thickness of the cylindrical reaction vessel is less than 1 mm, preferably less than 0.5 mm, in particular less than 0.3 mm. As a result, good heat transfer to the interior of the reaction vessel is possible. The heat transfer is better, the thinner the wall thickness. Most preferably, the wall thickness is as low as it can just be made by an injection molding process.
  • the liquid-conducting system or the opening or the first liquid line between the steps lit. a) and lit. b) is sealed liquid-tight. It is understood that the opening or the first liquid line after step lit. b) is opened again when the first liquid is then to be passed out of the reaction vessel again. By closing the liquid-conducting system, after the introduction of the first liquid any contamination of the environment can be avoided. This is especially important for infectious samples. It is very particularly preferred if the nucleic acid, in particular in the form of a biological sample, before step lit. a) is introduced into the liquid-conducting system and the liquid-conducting system after and before performing the step lit. a) is sealed liquid-tight.
  • Both the NSV and the detection can be done within the liquid-tight liquid-conducting system. Thereafter, the liquid-conducting system, including any liquids contained therein, can be disposed of completely. As a result, the inventive method is very safe. By closing the opening or the first liquid line, it can be achieved that a pressure building up in the reaction vessel can not affect the remaining liquid-conducting system.
  • the reaction vessel is lit. b) to set the conditions required by the NSA, in particular by blowing with an air stream, heated or cooled. Due to the cylindrical shape of the reaction vessel, the reaction vessel can be isolated and exposed so that substantially only the reaction vessel is heated or cooled. By blowing with a heated or cooled air flow, a particularly rapid temperature change within the reaction vessel and thereby a quick execution of the NSV is possible.
  • the reaction vessel in step lit. b) a cyclic temperature profile. This means that several times the same temperature levels are passed through within a given time sequence. It is particularly preferred if the
  • Air flow through an air duct, in particular an air deflector, at least partially, is guided around the reaction vessel.
  • the proof according to step lit. c) takes place outside the reaction vessel and the first liquid between the steps lit. b) and lit. c) by the opening and the first liquid line is passed, in particular by advancing the first piston, out of the reaction vessel.
  • the detection can be effected, for example, by means of an electrode with capture molecules immobilized thereon, with which the amplified nucleic acids hybridize, by means of an electrochemical reaction.
  • the first liquid is preferably conducted via the liquid-conducting system into a detection chamber in fluid communication with the liquid-conducting system.
  • a hybridization of the amplified nucleic acids with, in particular immobilized, further nucleic acids.
  • the further nucleic acids can be immobilized on a chip, in particular arranged in the detection chamber.
  • the proof according to step lit. c) can be done on the chip by means of an optical or an electrical detection method.
  • the presence of nucleic acids which have bound to other nucleic acids immobilized on the chip can be detected, for example, by measuring the fluorescence of fluorophores incorporated into the amplified nucleic acids.
  • the electrochemical detection can be carried out, for example, by means of graphic electrodes arranged on the chip, to which the further nucleic acids are immobilized as probes. After binding of the specific amplified nucleic acids, electrochemical detection of bound nucleic acids can then be performed.
  • an injection molded Plastic preferably polycarbonate
  • the liquid-conducting system comprises a line plate containing at least partially open lines and a closure means connected thereto, which closes the open lines.
  • the closing means can z.
  • Such a liquid-conducting system is simple and inexpensive to manufacture and has a high density and pressure stability of the lines.
  • the reaction vessel can also be provided in the form of a cartridge, which before the step lit. a) is connected in liquid-tight manner by plugging onto a receiving unit provided for this purpose and fluid-tightly to the outside with the liquid-conducting system.
  • the receiving unit can be provided for example by a connecting piece for inserting a connecting portion of the cartridge.
  • At least one cylindrical vessel which contains a second or further liquid and is connected fluid-conducting to the liquid-conducting system, the vessel having between an outer space and an inner space of the vessel a second or further piston displaceable in the vessel, wherein the second or further liquid is pressed into the reaction vessel by actuating the second or further piston via the liquid-conducting system. It is preferred if three cylindrical vessel is provided, which contains a second or further liquid and is connected fluid-conducting to the liquid-conducting system, the vessel having between an outer space and an inner space of the vessel a second or further piston displaceable in the vessel, wherein the second or further liquid is pressed into the reaction vessel by actuating the second or further piston via the liquid-conducting system. It is preferred if three cylindrical vessel is provided, which contains a second or further liquid and is connected fluid-conducting to the liquid-conducting system, the vessel having between an outer space and an inner space of the vessel a second or further piston displaceable in the vessel, wherein the second or further liquid
  • Containers are provided, each having a liquid and a flask, each containing one of the liquids required for performing a PCR reagent, in particular polymerase, primer and buffer.
  • the cylindrical vessel is provided in the form of a cartridge, which before the step lit. a) by connecting to a dedicated receiving unit with the liquid-conducting system liquid-conducting and liquid-tightly connected to the outside.
  • a dedicated receiving unit with the liquid-conducting system liquid-conducting and liquid-tightly connected to the outside.
  • the cylindrical vessel or, in the case of the provision of a plurality of cylindrical vessels the cylindrical vessels contain or contain all the reagents required for carrying out the NSV in addition to the nucleic acid, in particular polymerase, primer and buffer.
  • the liquid-conducting system comprises a plurality of second liquid lines arranged in a plane in a plate-like conduit element and the receiving unit / receiving units for attaching the cartridge / cartridges.
  • the implementation of the method with such a liquid-conducting system is particularly advantageous because the specificity of the method for certain nucleic acids can be chosen arbitrarily by attaching appropriate cartridges.
  • the second fluid lines can be provided, in particular provided by a membrane, valves.
  • the membrane is arranged and designed so that by applying pressure, a passage through the second liquid lines can be shut off.
  • the pressure may, for example, be exerted by a plunger actuated by an automated actuator on the diaphragm.
  • the liquid-conducting system may have a respective sample preparation chamber and waste chamber in fluid communication with the liquid-conducting system, as well as a connection pipe in fluid communication with the liquid-conducting system, in particular with a profile for producing a bayonet closure.
  • a tube containing a biological sample such as a so-called Monovette TM
  • the sample preparation chambers, the waste chamber, the connecting piece and the liquid-conducting system are integrally made of the same material, in particular of injection-molded plastic, preferably polycarbonate. It does not conflict with the one-piece construction if the conduit element comprises a conduit plate containing at least partially open conduits and a closure means connected thereto and closing off the open conduits.
  • the first and the second piston or, in the case of providing a plurality of cylindrical vessels, the first, the second and the further piston (s) is actuated by feed from the same side of the plate-like conduit element.
  • the invention further relates to an apparatus for carrying out the method according to the invention, consisting of a cylindrical reaction vessel, which is connected in liquid-conducting manner via a provided at a first end of the reaction vessel opening and opening into the opening first liquid line to a liquid-conducting system, so that the reaction vessel via the liquid-conducting system can be filled and emptied with a first, second or further liquid.
  • the reaction vessel has, between an outer space and an inner space of the reaction vessel, a first piston which is displaceable in the reaction vessel, wherein the first piston is designed so that it can be moved through the first, second or further liquid as it flows through the reaction vessel Opening is introduced or changes its volume or gas forms at a temperature change, thereby changing the volume of the interior of the reaction vessel is changed.
  • At least one vessel is provided, which contains the second or further liquid and is connected in a fluid-conducting manner to the liquid-conducting system.
  • the vessel or, in the case of the presence of multiple vessels, the vessels contain / contain reagents necessary for the performance of the NSV in addition to the nucleic acid. All can be included for performing the NSV in addition to the nucleic acid to be amplified reagents. If some of the reagents required for NSA are already introduced into the reaction vessel or contained in the first fluid, only the remaining part of the reagents necessary for the NSV can also be present.
  • the reaction vessel preferably has a rigid vessel wall, in particular consisting of polycarbonate.
  • a rigid vessel wall improves the tightness between the piston and the vessel wall.
  • Polycarbonate allows good heat transfer from the outside to the reaction vessel and is also compatible with performing a PCR.
  • the rigid vessel wall ensures a precise change in the volume of the interior of the reaction vessel and a precise
  • 1 is a schematic representation of a device according to the invention prior to carrying out the method according to the invention
  • 2 is a schematic representation of a device according to the invention at the beginning of filling the reaction vessel with a liquid
  • FIG. 3 is a schematic representation of a device according to the invention after filling with the liquid
  • Fig. 4 is a schematic representation of a device according to the invention in carrying out the method according to the invention.
  • Fig. 5 is an exploded view of a device according to the invention contained liquid-conducting system.
  • the device 10 consists of the reaction vessel 12 and a liquid-conducting system 14, to which the reaction vessel 12 via at least a first
  • the device 10 has a movable closure in the form of a first piston 18. Furthermore, the device 10 has a stop 21 formed by a removable rod 20 for the first piston 18.
  • the liquid-conducting system 14 has a closable or self-closing, designed here as a valve 22, inlet opening. Instead of the valve 22, a septum or a rubber stopper could also be provided here, which can be pierced by means of a cannula. Furthermore, the device 10 has a further valve 24.
  • Fig. 2 the beginning of the filling of the reaction vessel 12 with the first liquid 26 is shown schematically.
  • the The first liquid 26 is initially in the syringe 28.
  • the first liquid 26 is injected by means of a cannula through the open valve 22 into the liquid-conducting system 14. Since the further valve 24 is closed, the first liquid 26 is pressed into the reaction vessel 12 via the first liquid line 16. In this case, the first piston 18 is raised.
  • Into the reaction vessel 12 passes a precisely defined volume, which is predetermined by the stop 21 formed by the rod 20 for the first piston 18.
  • the syringe 28 is removed and the valve 22 is brought into the closed position. This condition is shown in FIG.
  • Fig. 4 shows the device 10 schematically in carrying out the method according to the invention.
  • the rod 20 has been removed to allow the first piston 18 to move freely.
  • the first liquid 26 is, as usual in performing a PCR, heated and cooled again. In this case, the volume of the first liquid changes 26. This change in volume causes a movement of the first piston 18. As a result, the volume change of the first liquid 26 is compensated by a corresponding change in volume of the interior of the reaction vessel 12. It is also possible to withdraw the rod 20 only slightly, so that by a movement of the first
  • Piston 18 Although the volume change of the first liquid 26 is indeed compensated, the formation of gas bubbles by a still existing formed by the rod 20 stop 21 for the first piston 18 is avoided or at least weakened.
  • FIG. 5 shows a fluidic system comprising a carrier 30. Liquid lines run on the underside of the carrier 30, which is not shown in the drawing. Finally, the first fluid line 16, which have a downwardly at least partially open side and are closed by the closure plate 32. On the support 30, the reaction vessel 12 is provided. For connecting cartridges 34 containing reagents further
  • connection element 38 is provided.
  • the connecting element 38 is connected to the carrier 30 and the chip 40, which serves to detect nucleic acids amplified in the reaction vessel 12, is latched to the latching hooks 42.
  • the liquid-conducting system 14 shown in Fig. 5 may form a unit after plugging the cartridges 34, in the complex processes such. For example, an opening of cells, a purification of biomolecules, a nucleic acid amplification and a detection of biomolecules can take place by means of the chip 40.
  • reaction vessel 14 liquid-conducting system

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  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Clinical Laboratory Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • Analytical Chemistry (AREA)
  • Hematology (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

L'invention concerne un procédé de réalisation d'une réaction de multiplication d'un acide nucléique (réaction de multiplication d'acide nucléique ; MAN) dans un système conduisant les liquides (14), l'acide nucléique étant contenu dans un premier liquide (26) et il est prévu un réacteur cylindrique (12) qui présente à une première extrémité un orifice qui est en communication fluide par le biais d'une première conduite de liquide (16) avec le système conduisant les liquides (14) et qui peut être rempli avec le premier liquide (26) et vidé par le biais de la première conduite de liquide (16), le réacteur (12) présentant entre un espace externe et l'espace interne du réacteur (12) un premier piston (18) mobile en translation dans le réacteur (12).
EP07711807A 2006-03-06 2007-03-06 Procédé de réalisation d'une réaction de multiplication d'un acide nucléique Withdrawn EP1993730A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE200610010958 DE102006010958A1 (de) 2006-03-06 2006-03-06 Verfahren zur Durchführung einer Reaktion zur Vervielfältigung einer Nukleinsäure
PCT/EP2007/001903 WO2007101653A1 (fr) 2006-03-06 2007-03-06 Procédé de réalisation d'une réaction de multiplication d'un acide nucléique

Publications (1)

Publication Number Publication Date
EP1993730A1 true EP1993730A1 (fr) 2008-11-26

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EP07711807A Withdrawn EP1993730A1 (fr) 2006-03-06 2007-03-06 Procédé de réalisation d'une réaction de multiplication d'un acide nucléique

Country Status (3)

Country Link
EP (1) EP1993730A1 (fr)
DE (1) DE102006010958A1 (fr)
WO (1) WO2007101653A1 (fr)

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Publication number Priority date Publication date Assignee Title
CN111205971B (zh) * 2020-04-22 2020-07-31 博奥生物集团有限公司 采样管

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CA1338505C (fr) * 1989-02-03 1996-08-06 John Bruce Findlay Cuvette de retenue pour pcr et methode d'utilisation
US6607907B2 (en) * 2000-05-15 2003-08-19 Biomicro Systems, Inc. Air flow regulation in microfluidic circuits for pressure control and gaseous exchange
DE10319045A1 (de) * 2003-04-25 2004-12-09 november Aktiengesellschaft Gesellschaft für Molekulare Medizin Vorrichtung und Verfahren zur Aufbereitung Biopolymerhaltiger Flüssigkeiten
EP1641564B1 (fr) * 2003-07-04 2007-10-31 november Aktiengesellschaft Gesellschaft für Molekulare Medizin Utilisation d'un contenant a usage unique, dispositif microfluidique et procede de traitement de molecules
DE202004012163U1 (de) * 2004-07-08 2004-10-07 Tecan Trading Ag System mit Einrichtung zum Verhindern von Luftblasen in einer Hybridisierkammer
DE102004058828B4 (de) * 2004-10-28 2010-08-19 Progen Biotechnik Gmbh Vorrichtung und Verfahren zur parallelen Aufbereitung von Biopolymeren

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Title
See also references of WO2007101653A1 *
WITTWER C.T. ET AL: "Minimizing the time required for DNA amplification by efficient heat transfer to small samples", ANALYTICAL BIOCHEMISTRY, ACADEMIC PRESS INC, NEW YORK, vol. 186, no. 2, 1 May 1990 (1990-05-01), pages 328 - 331, XP024823234, ISSN: 0003-2697, [retrieved on 19900501], DOI: DOI:10.1016/0003-2697(90)90090-V *

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Publication number Publication date
DE102006010958A1 (de) 2007-09-13
WO2007101653A1 (fr) 2007-09-13

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