FR2793260A1 - Device for grinding food sample to extract DNA, RNA, or proteins, has rotatable pestles carried on a block above a tray moving between working and retracted positions - Google Patents

Device for grinding food sample to extract DNA, RNA, or proteins, has rotatable pestles carried on a block above a tray moving between working and retracted positions Download PDF

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Publication number
FR2793260A1
FR2793260A1 FR9905795A FR9905795A FR2793260A1 FR 2793260 A1 FR2793260 A1 FR 2793260A1 FR 9905795 A FR9905795 A FR 9905795A FR 9905795 A FR9905795 A FR 9905795A FR 2793260 A1 FR2793260 A1 FR 2793260A1
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Prior art keywords
proteins
rna
dna
liquid
biological samples
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FR9905795A
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French (fr)
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FR2793260B1 (en
Inventor
Michel Gazeau
Vladimir Lazar
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Genomic SA
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Genomic SA
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Priority to FR9905795A priority Critical patent/FR2793260B1/en
Priority to FR9910040A priority patent/FR2793261B1/en
Priority to AU44141/00A priority patent/AU4414100A/en
Priority to AT00925408T priority patent/ATE279510T1/en
Priority to EP00925408A priority patent/EP1177276B1/en
Priority to PCT/FR2000/001230 priority patent/WO2000068358A1/en
Priority to DE2000614886 priority patent/DE60014886D1/en
Publication of FR2793260A1 publication Critical patent/FR2793260A1/en
Priority to US10/007,043 priority patent/US6695236B2/en
Application granted granted Critical
Publication of FR2793260B1 publication Critical patent/FR2793260B1/en
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M47/00—Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass
    • C12M47/06—Hydrolysis; Cell lysis; Extraction of intracellular or cell wall material

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  • Engineering & Computer Science (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Organic Chemistry (AREA)
  • Biomedical Technology (AREA)
  • Genetics & Genomics (AREA)
  • Biotechnology (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Microbiology (AREA)
  • Molecular Biology (AREA)
  • Cell Biology (AREA)
  • Sustainable Development (AREA)
  • Analytical Chemistry (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Biophysics (AREA)
  • Plant Pathology (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Sampling And Sample Adjustment (AREA)

Abstract

A device for grinding a biological sample comprising rotatable pestles each having a liquid supply conduit, is new. The pestles are carried on a block above a tray that moves between a working position adjacent to the pestles and a retracted position.

Description

Procédé et dispositif pour l'extraction d'ADN, d'ARN et de protéines àMethod and device for the extraction of DNA, RNA and proteins from

partir de prélèvements biologiques non liquides La présente invention concerne le domaine de l'extraction d'ADN, d'ARN et de protéines à haut débit, à partir de prélèvements biologiques non liquides, à l'état naturel ou entrant dans des préparations par exemple alimentaires. On connaît dans l'état de la technique différents procédés d'extraction d'acides nucléiques et de protéines à partir de prélèvements non liquides mettent en jeu une première étape de broyage de l'échantillon, par des procédés  The present invention relates to the field of high-throughput DNA, RNA and protein extraction from non-liquid biological samples, in their natural state or entering into preparations for example. food. Various methods of extracting nucleic acids and proteins from non-liquid samples are known in the state of the art, involving a first step of grinding the sample, by methods

divers tels pilage, ultrasons, haute pression...  various such pounding, ultrasound, high pressure ...

Il est intéressant, pour des échantillons mous, de les broyer à très basse température. En effet, cela diminue les risques de dégradation des macromolécules et facilite l'éclatement des cellules et donc améliore le rendement des extractions. Il s'en suit différents traitements de l'échantillon, qui visent à le laver, à supprimer les  It is interesting, for soft samples, to grind them at very low temperature. Indeed, this reduces the risks of degradation of the macromolecules and facilitates the bursting of the cells and therefore improves the yield of the extractions. It follows different treatments of the sample, which aim to wash it, to remove the

membranes cellulaires et à éliminer les produits non voulus.  cell membranes and to remove unwanted products.

La première étape n'étant pas systématiquement mise en oeuvre.  The first step is not systematically implemented.

Le procédé faisant l'objet de la demande de brevet comporte les étapes suivantes: Il consiste, après broyage à basse température, (température inférieure à la température ambiante) par des procédés divers, simultanément, d'un nombre d'échantillons, à ajouter automatiquement et avec un temps d'attente minimal, un volume suffisamment précis de liquide/s, à chaque échantillon broyé, tout en contrôlant la température du broyat, du/des liquide/s et du mélange et à homogénéiser  The process which is the subject of the patent application comprises the following stages: It consists, after grinding at low temperature (temperature below ambient temperature) by various processes, simultaneously, of a number of samples, to be added automatically and with a minimum waiting time, a sufficiently precise volume of liquid / s, for each ground sample, while controlling the temperature of the ground material, of the liquid / s and of the mixture and to homogenize

chaque mélange.each mixture.

L'un des liquides peut de préférence être un tampon de lyse des parois cellulaires et plus précisément, un tampon  One of the liquids may preferably be a cell wall lysis buffer and more specifically, a buffer

de lyse dénaturant.denaturing lysis.

2 27932602 2793260

Il s'agit en particulier d'éviter que les agents de dégradation des macro-molécules puissent agir, tout en permettant l'écoulement du/des liquide/s, même si le broyage a eu lieu à très basse température (inférieure à 0 C) Il s'agit de limiter les risques de contamination des personnes, par des virus, par exemple et de diminuer la perte de matériel biologique qui adhérerait aux pièces de broyage. Il permet le traitement plusieurs échantillons simultanément, et l'enchaînement des opérations et leur automatisation. Le dispositif préféré, décrit à titre d'exemple est constitué des éléments suivants: * L'équipement de broyage de base, qui est dédié aux échantillons végétaux, pour lesquels seules les protéines  This is in particular to avoid that the agents of degradation of the macro-molecules can act, while allowing the flow of the liquid (s), even if the grinding took place at very low temperature (below 0 C ) It is a question of limiting the risks of contamination of the people, by viruses, for example and of decreasing the loss of biological material which would adhere to the pieces of grinding. It allows the processing of several samples simultaneously, and the sequence of operations and their automation. The preferred device, described by way of example, consists of the following elements: * The basic grinding equipment, which is dedicated to plant samples, for which only the proteins

comptent, (Voir description du Equipement de broyage en  count, (See description of the Grinding Equipment in

annexe) auquel est ajouté: * Un dispositif constitué d'un réseau de buses, chacune étant située au voisinage de la tige de chaque pilon, communicant par des cathéters avec le/les réservoir/s du/des liquide/s à distribuer. Les réservoir peuvent être maintenus à des températures définies. Le/les liquides peuvent s'écouler par gravité, lors de l'ouverture d'électrovannes et le volume distribué réglé grâce à des cathéters de longueur et de diamètre précis, en jouant sur la durée d'ouverture des électrovannes. Une autre solution est par exemple d'aspirer et de refouler le/les liquides au moyen de pompes de  appendix) to which is added: * A device consisting of a network of nozzles, each being located in the vicinity of the rod of each pestle, communicating by catheters with the reservoir (s) of the liquid (s) to be dispensed. The tanks can be maintained at set temperatures. The liquid (s) can flow by gravity when the solenoid valves are opened and the dispensed volume adjusted using catheters of precise length and diameter, by varying the duration of the solenoid valve opening. Another solution is, for example, to suck and discharge the liquid (s) by means of

précision. Dans ce cas il est facile de purger les cathéters.  precision. In this case it is easy to purge the catheters.

Les tiges de pilons servent de guide à l'écoulement du/des  The drumstick rods serve as a guide for the flow of the

liquides vers les échantillons.liquids to samples.

* Des dispositifs de chauffage régulés, qui peuvent être des résistances noyées dans le plateau porte tubes du Équipement de broyage et/ou des rampes à infrarouge disposées  * Regulated heating devices, which can be resistors embedded in the tube-holder tray of the grinding equipment and / or infrared ramps arranged

autour de l'espace de travail.around the workspace.

* Accessoirement, une plaque rigide, traversée par les tiges des pilons peut être rajoutée au dispositif préféré, pour couvrir chaque échantillon dans son conteneur, pendant le broyage, afin d'éviter des contaminations croisées. Cette plaque, grâce aux fraisages de chaque passage de tige de pilon sur la partie supérieure, favorise l'écoulement uniforme du/des liquides dans chaque conteneur d'échantillon, autour des pilons  * Incidentally, a rigid plate, crossed by the rods of the drumsticks can be added to the preferred device, to cover each sample in its container, during grinding, in order to avoid cross contamination. This plate, thanks to the milling of each passage of the pestle rod on the upper part, promotes the uniform flow of the liquid (s) in each sample container, around the pestles

et contribue à les débarrasser des fragments adhérents.  and helps to rid them of adherent fragments.

L'automate programmable qui contrôle l'équipement de broyage contrôle également les écoulements de liquide, les  The programmable controller that controls the grinding equipment also controls the liquid flows,

températures et le verrouillage de la plaque rigide.  temperatures and locking the rigid plate.

La présente invention concerne en particulier un dispositif pour l'extraction d'ADN, d'ARN et de protéines à partir de prélèvements biologiques non liquides, caractérisé en ce qu'il comprend en outre des moyens pour permettre l'écoulement du ou des liquides par gravité, lors de  The present invention relates in particular to a device for the extraction of DNA, RNA and proteins from non-liquid biological samples, characterized in that it further comprises means for allowing the flow of the liquid (s) by gravity, when

l'ouverture d'électrovannes.opening of solenoid valves.

4 27932604 2793260

Claims (6)

REVENDICATIONS 1 - Procédé d'extraction d'ADN, d'ARN et de protéines à partir de prélèvements biologiques non liquides caractérisé en ce qu'il comporte les étapes suivantes: - broyage à température inférieure à la température ambiante - par des procédés divers, simultanément, d'un nombre d'échantillons, - ajout automatique d'au moins un liquide à chaque échantillon broyé, - contrôle de la température du broyat, du/des  1 - Method for extracting DNA, RNA and proteins from non-liquid biological samples, characterized in that it comprises the following stages: - grinding at a temperature below room temperature - by various methods, simultaneously , of a number of samples, - automatic addition of at least one liquid to each ground sample, - control of the temperature of the ground material, liquide/s et du mélange et à homogénéiser chaque mélange.  liquid / s and mixture and homogenize each mixture. 2 - Procédé d'extraction d'ADN, d'ARN et de protéines à partir de prélèvements biologiques non liquides selon la revendication 1 caractérisé en ce qu'un des liquides est constitué par un tampon de lyse des parois cellulaires et  2 - Method for extracting DNA, RNA and proteins from non-liquid biological samples according to claim 1 characterized in that one of the liquids consists of a lysis buffer for the cell walls and plus précisément, un tampon de lyse dénaturant.  more specifically, a denaturing lysis buffer. 3 - Dispositif pour l'extraction d'ADN, d'ARN et de protéines à partir de prélèvements biologiques non liquides préféré, caractérisé en ce qu'il comprend * un équipement de broyage de base, qui est dédié aux échantillons végétaux, pour lesquels seules les protéines comptent, * Un dispositif constitué d'un réseau de buses, chacune étant située au voisinage de la tige de chaque pilon, communicant par des cathéters avec le/les réservoir/s du/des liquide/s à distribuer. Les réservoir peuvent être maintenus  3 - Device for the extraction of DNA, RNA and proteins from preferred non-liquid biological samples, characterized in that it comprises * basic grinding equipment, which is dedicated to plant samples, for which only proteins count, * A device made up of a network of nozzles, each being located in the vicinity of the rod of each pestle, communicating by catheters with the reservoir (s) of the liquid (s) to be dispensed. Tanks can be maintained à des températures définies.at defined temperatures. 4 - Dispositif pour l'extraction d'ADN, d'ARN et de protéines à partir de prélèvements biologiques non liquides selon la revendication 3, caractérisé en ce qu'il comprend en outre des moyens pour permettre l'écoulement du ou des  4 - Device for the extraction of DNA, RNA and proteins from non-liquid biological samples according to claim 3, characterized in that it further comprises means for allowing the flow of the or liquides par gravité, lors de l'ouverture d'électrovannes.  liquids by gravity when opening solenoid valves. - Dispositif pour l'extraction d'ADN, d'ARN et de protéines à partir de prélèvements biologiques non liquides selon la revendication 3, caractérisé en ce qu'il comprend des  - Device for the extraction of DNA, RNA and proteins from non-liquid biological samples according to claim 3, characterized in that it comprises pompes et des moyens de purge des cathéters.  pumps and means for purging catheters. 6 - Dispositif pour l'extraction d'ADN, d'ARN et de protéines à partir de prélèvements biologiques non liquides  6 - Device for the extraction of DNA, RNA and proteins from non-liquid biological samples selon la revendication 3, caractérisé en ce qu'il en outre des dispositifs de chauffage régulés.  according to claim 3, characterized in that it furthermore regulated heating devices. 7 - Dispositif pour l'extraction d'ADN, d'ARN et de protéines à partir de prélèvements biologiques non liquides selon la revendication 3, caractérisé en ce qu'il en outre une0 plaque rigide, traversée par les tiges des pilons peut être rajoutée au dispositif préféré, pour couvrir chaque échantillon  7 - Device for the extraction of DNA, RNA and proteins from non-liquid biological samples according to claim 3, characterized in that it furthermore a rigid plate, crossed by the rods of the drumsticks can be added to the preferred device, to cover each sample dans son conteneur, pendant le broyage, afin d'éviter des contaminations croisées.  in its container, during grinding, in order to avoid cross contamination.
FR9905795A 1999-05-05 1999-05-05 METHOD AND DEVICE FOR THE EXTRACTION OF DNA, RNA AND PROTEINS FROM NON-LIQUID BIOLOGICAL SAMPLES Expired - Fee Related FR2793260B1 (en)

Priority Applications (8)

Application Number Priority Date Filing Date Title
FR9905795A FR2793260B1 (en) 1999-05-05 1999-05-05 METHOD AND DEVICE FOR THE EXTRACTION OF DNA, RNA AND PROTEINS FROM NON-LIQUID BIOLOGICAL SAMPLES
FR9910040A FR2793261B1 (en) 1999-05-05 1999-08-02 PROCESS AND DEVICE FOR THE EXTRACTION OF DNA, RNA AND PROTEINS FROM BIOLOGICAL SAMPLES
AT00925408T ATE279510T1 (en) 1999-05-05 2000-05-05 DEVICE FOR OBTAINING DNA, RNA AND PROTEINS FROM BIOLOGICAL SAMPLES
EP00925408A EP1177276B1 (en) 1999-05-05 2000-05-05 Device for extracting DNA, RNA and proteins from biological samples
AU44141/00A AU4414100A (en) 1999-05-05 2000-05-05 Method and device for extracting dna, rna and proteins from biological samples
PCT/FR2000/001230 WO2000068358A1 (en) 1999-05-05 2000-05-05 Method and device for extracting dna, rna and proteins from biological samples
DE2000614886 DE60014886D1 (en) 1999-05-05 2000-05-05 Device for obtaining DNA, RNA and proteins from biological samples
US10/007,043 US6695236B2 (en) 1999-05-05 2001-11-05 Process and device for the extraction of DNA, RNA and proteins from biological samples

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FR9905795A FR2793260B1 (en) 1999-05-05 1999-05-05 METHOD AND DEVICE FOR THE EXTRACTION OF DNA, RNA AND PROTEINS FROM NON-LIQUID BIOLOGICAL SAMPLES

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FR2793260A1 true FR2793260A1 (en) 2000-11-10
FR2793260B1 FR2793260B1 (en) 2003-05-16

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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0487028A2 (en) * 1990-11-22 1992-05-27 Shimadzu Corporation Apparatus for extracting and purifying nucleic acid
US5330916A (en) * 1990-06-26 1994-07-19 E. I. Du Pont De Nemours And Company Cellular component extraction apparatus and disposable vessel useful therein
WO1998020164A1 (en) * 1996-11-04 1998-05-14 Cornell Research Foundation, Inc. Matrix mill for dna extraction

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5330916A (en) * 1990-06-26 1994-07-19 E. I. Du Pont De Nemours And Company Cellular component extraction apparatus and disposable vessel useful therein
EP0487028A2 (en) * 1990-11-22 1992-05-27 Shimadzu Corporation Apparatus for extracting and purifying nucleic acid
WO1998020164A1 (en) * 1996-11-04 1998-05-14 Cornell Research Foundation, Inc. Matrix mill for dna extraction

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