WO2004108925A1 - Methode d'isolation sequentielle d'adn et d'arn a partir du meme echantillon contenant des acides nucleiques - Google Patents

Methode d'isolation sequentielle d'adn et d'arn a partir du meme echantillon contenant des acides nucleiques Download PDF

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Publication number
WO2004108925A1
WO2004108925A1 PCT/EP2004/005999 EP2004005999W WO2004108925A1 WO 2004108925 A1 WO2004108925 A1 WO 2004108925A1 EP 2004005999 W EP2004005999 W EP 2004005999W WO 2004108925 A1 WO2004108925 A1 WO 2004108925A1
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WIPO (PCT)
Prior art keywords
functional surface
dna
solution
rna
time period
Prior art date
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PCT/EP2004/005999
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English (en)
Inventor
Evy H. Reitan
Arne Deggerdal
Vidar Skagestad
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Qiagen As
Qiagen 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.)
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Publication date
Application filed by Qiagen As, Qiagen Gmbh filed Critical Qiagen As
Priority to EP04739565A priority Critical patent/EP1633869A1/fr
Publication of WO2004108925A1 publication Critical patent/WO2004108925A1/fr

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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
    • 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
    • C12N15/1006—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers

Definitions

  • the present invention refers to a method for isolating nucleic acids from a nucleic acid-containing sample. More specifically, the present invention relates to a method for sequentially isolating DNA and RNA from the same nucleic acid-containing sample. Furthermore, the present invention refers to kit for sequentially isolating DNA and RNA from the same nucleic acid-containing sample.
  • RNA isolation procedures show some degree of selectivity between DNA and RNA.
  • the selective isolation of RNA is performed either under acidic conditions on a silica solid phase (US 5,990,302) or under chaotropic conditions in the presence of an alcohol on a silica solid phase (WO 95/01359). In the latter case, normally a low selectivity for RNA over DNA is observed, and a DNA digest step is thus included to obtain pure RNA.
  • some methods for the selective isolation of DNA are known from the art.
  • step (a) adding a chaotropic salt to a final concentration of from 1 to 4.5 M to a nucleic acid-containing sample, (b) following step (a), adding an alcohol to a final concentration of from 13 to 70 %(v/v) to the solution of (a),
  • step (c) bringing the solution of step (b) into contact with a functional surface, maintaining this contact for a particular time period, breaking the contact between the solution of step (b) and the functional surface,
  • step (b) following step (a), adding an alcohol to a final concentration of from 13 to 70 %(v/v) to the solution of (a),
  • the functional surface utilized in step (d) is of the same kind as the functional surface of step (c).
  • the method according to the invention is, thereby, less laborious and easier to handle.
  • the binding conditions for DNA and RNA do not have to be changed between step (c) and step (d) due to the same kind of solid support utilized in step (c) and step (d).
  • a functional surface useful in the present invention is either a surface comprising carboxylic acid groups or, preferably, the functional surface is a silica surface.
  • the functional surface is provided in bead form. If the functional surface is provided in bead form, preferably the functional surface comprises a plurality of beads.
  • the chaotropic salt is added in step (a) in form of a solution and can advantageously be used to lyse the source of the nucleic acids, e.g. cells or tissue. In this case an additional sufficient incubation time is needed to allow the cells to lyse.
  • the required conditions to lyse the source of the nucleic acids i.e. incubation time, temperature etc., are well known to a person skilled in the art and can easily be adapted to the method according to the invention.
  • the chaotropic salt is added to the nucleic acid-containing sample as a solution of suitable concentration.
  • suitable solvent e.g. water or a buffer system
  • Suitable solvents or buffer systems according to the present invention are obvious to a skilled person.
  • the chaotropic salt can be added as a solid. In the latter case, it is required that the nucleic acid-containing sample is available as a solution.
  • the term 'final concentration' for the purpose of the present invention stands for the concentration of a substance, i.e. the chaotropic salt added in step (a) and the alcohol added in step (b), after adding the chaotropic salt in step (a) and after adding the alcohol in step (b) but before step (c) of the present invention.
  • the alcohol added in step (b) is selected from methanol, ethanol, n-propanol, iso- propanol or is a mixture thereof. Thereby, the alcohol is added pure or diluted in a suitable solvent, e.g. water, to a suitable concentration.
  • a suitable solvent e.g. water
  • step (c) may comprise at least the steps of:
  • step (3) prior to, simultaneously with or following step (1) or step (2), performing a
  • a suitable solution comprising either a high concentration of chaotropic salt, e.g. 7 M guanidinium hydrochloride, or a high concentration of a suitable alcohol, e.g. ethanol at 65 to 80 % (v/v), or a suitable organic solvent, the nucleic acids being insoluble in this organic solvent.
  • a suitable solution compositions and performance of such washing steps are known from the state of the art. At least one washing step may be performed or the washing steps may be performed in a number seeming suitable to a person skilled in the art.
  • GtB-CtA concentration B] E " .
  • step (a) 6 mg of ferrimagnetic particles were suspended in 20 ⁇ of an aqueous solution comprising a composition according to the final composition of the lysate as displayed above (68.6 %(v/v) ethanol, 1.0 M guanidinium thiocyanate, 7.2 mM trisodium citrate).

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  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Health & Medical Sciences (AREA)
  • Biomedical Technology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Plant Pathology (AREA)
  • Molecular Biology (AREA)
  • Microbiology (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

La présente invention concerne une méthode permettant d'isoler des acides nucléiques à partir d'un échantillon contenant des acides nucléiques. Plus spécifiquement, ladite invention a trait à une méthode permettant d'isoler séquentiellement l'ADN et l'ARN à partir du même échantillon contenant des acides nucléiques. En outre, cette invention a trait à un kit destiné à l'isolation séquentielle d'ADN et d'ARN à partir du même échantillon contenant des acides nucléiques.
PCT/EP2004/005999 2003-06-04 2004-06-03 Methode d'isolation sequentielle d'adn et d'arn a partir du meme echantillon contenant des acides nucleiques WO2004108925A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
EP04739565A EP1633869A1 (fr) 2003-06-04 2004-06-03 Methode d'isolation sequentielle d'adn et d'arn a partir du meme echantillon contenant des acides nucleiques

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US47607203P 2003-06-04 2003-06-04
US60/476,072 2003-06-04

Publications (1)

Publication Number Publication Date
WO2004108925A1 true WO2004108925A1 (fr) 2004-12-16

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PCT/EP2004/005999 WO2004108925A1 (fr) 2003-06-04 2004-06-03 Methode d'isolation sequentielle d'adn et d'arn a partir du meme echantillon contenant des acides nucleiques

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EP (1) EP1633869A1 (fr)
WO (1) WO2004108925A1 (fr)

Cited By (24)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1996730A2 (fr) * 2006-03-08 2008-12-03 Promega Corporation Purification de petits arn
WO2009040444A1 (fr) * 2007-09-28 2009-04-02 Mole Genetics As Procédé d'isolement d'arn
WO2009070465A1 (fr) * 2007-11-29 2009-06-04 New England Biolabs, Inc. Purification sélective de petits arn issus de mélanges
EP2264168A1 (fr) * 2009-06-18 2010-12-22 Qiagen GmbH Procédé d'isolation d'acides nucléiques
US8030034B2 (en) 2005-12-09 2011-10-04 Promega Corporation Nucleic acid purification with a binding matrix
WO2012002887A1 (fr) * 2010-06-29 2012-01-05 Sjoeblom Tobias Procédé et kit d'isolation séquentielle d'espèces de nucléotide provenant d'un échantillon
US20120288867A1 (en) * 2011-05-12 2012-11-15 Exact Sciences Corporation Serial isolation of multiple dna targets from stool
JP2013516969A (ja) * 2010-01-18 2013-05-16 キアゲン ゲーエムベーハー 小型ｒｎａを単離するための方法
US8980107B2 (en) 2011-05-12 2015-03-17 Exact Sciences Corporation Spin filter
US8993341B2 (en) 2011-05-12 2015-03-31 Exact Sciences Corporation Removal of PCR inhibitors
EP2285816B1 (fr) 2008-05-30 2015-04-01 Qiagen GmbH Procédé d'isolation d'acides nucléiques à chaînes courtes
US8999176B2 (en) 2011-05-12 2015-04-07 Exact Sciences Corporation Isolation of nucleic acids
WO2016077294A1 (fr) * 2014-11-14 2016-05-19 Corning Incorporated Procédés et kits pour la purification d'arn post-tiv
US9453257B2 (en) 2006-05-31 2016-09-27 Sequenom, Inc. Methods and compositions for the extraction and amplification of nucleic acid from a sample
WO2016198519A1 (fr) * 2015-06-09 2016-12-15 Biocartis N.V. Procédé automatisable pour l'isolement d'acides nucléiques
US9580741B2 (en) 2009-04-03 2017-02-28 Sequenom, Inc. Nucleic acid preparation compositions and methods
EP3138913A1 (fr) * 2015-09-02 2017-03-08 Imagen Bioscience Procédé et kit d'isolement sélectif d'acide nucléique
RU188425U1 (ru) * 2018-08-02 2019-04-11 Мераб Георгиевич Чикобава Дозированная форма для выделения днк
WO2019209597A1 (fr) 2018-04-24 2019-10-31 Qiagen Sciences Llc Isolement d'acide nucléique et élimination d'inhibiteur vis-à-vis d'échantillons complexes
WO2019207168A1 (fr) 2018-04-27 2019-10-31 Qiagen Gmbh Procédé d'isolement d'acides nucléiques à partir d'échantillons de plantes
WO2019209600A1 (fr) 2018-04-24 2019-10-31 Qiagen Sciences Llc Isolement de biomolécules et élimination d'inhibiteur
WO2019214988A1 (fr) 2018-05-11 2019-11-14 Qiagen Gmbh Procédé de lyse pour échantillons végétaux
US11242518B2 (en) 2015-09-04 2022-02-08 QIAGEN Sciences, LLP Methods for co-isolation of nucleic acids and proteins
US11827929B2 (en) 2015-07-23 2023-11-28 Biocartis, Nv Optimized clinical sample sequencing

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US8030034B2 (en) 2005-12-09 2011-10-04 Promega Corporation Nucleic acid purification with a binding matrix
EP1996730A2 (fr) * 2006-03-08 2008-12-03 Promega Corporation Purification de petits arn
EP1996730A4 (fr) * 2006-03-08 2010-03-03 Promega Corp Purification de petits arn
US9453257B2 (en) 2006-05-31 2016-09-27 Sequenom, Inc. Methods and compositions for the extraction and amplification of nucleic acid from a sample
US11952569B2 (en) 2006-05-31 2024-04-09 Sequenom, Inc. Methods and compositions for the extraction and amplification of nucleic acid from a sample
US10662421B2 (en) 2006-05-31 2020-05-26 Sequenom, Inc. Methods and compositions for the extraction and amplification of nucleic acid from a sample
GB2466419A (en) * 2007-09-28 2010-06-23 Mole Genetics As RNA isolation method
GB2466419B (en) * 2007-09-28 2011-04-13 Mole Genetics As RNA isolation method
WO2009040444A1 (fr) * 2007-09-28 2009-04-02 Mole Genetics As Procédé d'isolement d'arn
WO2009070465A1 (fr) * 2007-11-29 2009-06-04 New England Biolabs, Inc. Purification sélective de petits arn issus de mélanges
US10738069B2 (en) 2008-05-30 2020-08-11 Qiagen Gmbh Method for isolating nucleic acids
US9790250B2 (en) 2008-05-30 2017-10-17 Qiagen Gmbh Method for isolating short-chain nucleic acids
US9809612B2 (en) 2008-05-30 2017-11-07 Qiagen Gmbh Method for isolating nucleic acids
EP2285816B1 (fr) 2008-05-30 2015-04-01 Qiagen GmbH Procédé d'isolation d'acides nucléiques à chaînes courtes
US10053685B2 (en) 2009-04-03 2018-08-21 Sequenom, Inc. Nucleic acid preparation compositions and methods
US10858645B2 (en) 2009-04-03 2020-12-08 Sequenom, Inc. Nucleic acid preparation compositions and methods
US9580741B2 (en) 2009-04-03 2017-02-28 Sequenom, Inc. Nucleic acid preparation compositions and methods
US9850480B2 (en) 2009-04-03 2017-12-26 Sequenom, Inc. Nucleic acid preparation compositions and methods
US8980552B2 (en) 2009-06-18 2015-03-17 Qiagen Gmbh Method for isolating nucleic acids
WO2010145843A1 (fr) * 2009-06-18 2010-12-23 Qiagen Gmbh Procédé pour isoler des acides nucléiques
EP2264168A1 (fr) * 2009-06-18 2010-12-22 Qiagen GmbH Procédé d'isolation d'acides nucléiques
JP2013516969A (ja) * 2010-01-18 2013-05-16 キアゲン ゲーエムベーハー 小型ｒｎａを単離するための方法
US8889393B2 (en) 2010-06-29 2014-11-18 Exscale Biospecimen Solutions Ab Method and kit for sequential isolation of nucleotide species from a sample
WO2012002887A1 (fr) * 2010-06-29 2012-01-05 Sjoeblom Tobias Procédé et kit d'isolation séquentielle d'espèces de nucléotide provenant d'un échantillon
US9631228B2 (en) 2011-05-12 2017-04-25 Exact Sciences Corporation Isolation of nucleic acids
US8980107B2 (en) 2011-05-12 2015-03-17 Exact Sciences Corporation Spin filter
US20120288867A1 (en) * 2011-05-12 2012-11-15 Exact Sciences Corporation Serial isolation of multiple dna targets from stool
US11674169B2 (en) 2011-05-12 2023-06-13 Exact Sciences Corporation Isolation of nucleic acids
US9657330B2 (en) 2011-05-12 2017-05-23 Exact Sciences Corporation Serial isolation of multiple DNA targets from stool
US9163278B2 (en) 2011-05-12 2015-10-20 Exact Sciences Corporation Isolation of nucleic acids
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US9856515B2 (en) 2011-05-12 2018-01-02 Exact Sciences Corporation Removal of PCR inhibitors
US8808990B2 (en) * 2011-05-12 2014-08-19 Exact Sciences Corporation Serial isolation of multiple DNA targets from stool
US10047390B2 (en) 2011-05-12 2018-08-14 Exact Sciences Corporation Isolation of nucleic acids
US8999176B2 (en) 2011-05-12 2015-04-07 Exact Sciences Corporation Isolation of nucleic acids
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US8993341B2 (en) 2011-05-12 2015-03-31 Exact Sciences Corporation Removal of PCR inhibitors
WO2016077294A1 (fr) * 2014-11-14 2016-05-19 Corning Incorporated Procédés et kits pour la purification d'arn post-tiv
US10731147B2 (en) 2014-11-14 2020-08-04 Corning Incorporated Methods and kits for post-IVT RNA purification
US10364428B2 (en) 2014-11-14 2019-07-30 Corning Incorporated Methods and kits for post-IVT RNA purification
WO2016198519A1 (fr) * 2015-06-09 2016-12-15 Biocartis N.V. Procédé automatisable pour l'isolement d'acides nucléiques
US10829758B2 (en) 2015-06-09 2020-11-10 Biocartis, Nv Automatable method for nucleic acid isolation
CN107683335A (zh) * 2015-06-09 2018-02-09 拜奥卡蒂斯股份有限公司 用于核酸分离的自动化方法
EP4186980A1 (fr) * 2015-06-09 2023-05-31 Biocartis NV Procédé automatisable pour l'isolement d'acides nucléiques
US11827929B2 (en) 2015-07-23 2023-11-28 Biocartis, Nv Optimized clinical sample sequencing
EP3138913A1 (fr) * 2015-09-02 2017-03-08 Imagen Bioscience Procédé et kit d'isolement sélectif d'acide nucléique
US11242518B2 (en) 2015-09-04 2022-02-08 QIAGEN Sciences, LLP Methods for co-isolation of nucleic acids and proteins
US11814618B2 (en) 2015-09-04 2023-11-14 Qiagen Sciences, Llc Methods for co-isolation of nucleic acids and proteins
WO2019209600A1 (fr) 2018-04-24 2019-10-31 Qiagen Sciences Llc Isolement de biomolécules et élimination d'inhibiteur
WO2019209597A1 (fr) 2018-04-24 2019-10-31 Qiagen Sciences Llc Isolement d'acide nucléique et élimination d'inhibiteur vis-à-vis d'échantillons complexes
WO2019207168A1 (fr) 2018-04-27 2019-10-31 Qiagen Gmbh Procédé d'isolement d'acides nucléiques à partir d'échantillons de plantes
WO2019214988A1 (fr) 2018-05-11 2019-11-14 Qiagen Gmbh Procédé de lyse pour échantillons végétaux
RU188425U1 (ru) * 2018-08-02 2019-04-11 Мераб Георгиевич Чикобава Дозированная форма для выделения днк

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