WO2009129797A2 - Stabile lysepuffermixtur zur extraktion von nukleinsäuren - Google Patents
Stabile lysepuffermixtur zur extraktion von nukleinsäuren Download PDFInfo
- Publication number
- WO2009129797A2 WO2009129797A2 PCT/DE2009/000549 DE2009000549W WO2009129797A2 WO 2009129797 A2 WO2009129797 A2 WO 2009129797A2 DE 2009000549 W DE2009000549 W DE 2009000549W WO 2009129797 A2 WO2009129797 A2 WO 2009129797A2
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- lysis buffer
- buffer mixture
- nucleic acids
- mixture according
- extraction
- Prior art date
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Classifications
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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
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/96—Stabilising an enzyme by forming an adduct or a composition; Forming enzyme conjugates
-
- 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
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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
- 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
- C12N15/1013—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 by using magnetic beads
-
- 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
- C12N15/1017—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by filtration, e.g. using filters, frits, membranes
-
- 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
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
- C12N9/2462—Lysozyme (3.2.1.17)
-
- 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
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
- C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
- C12N9/64—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue
- C12N9/6421—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue from mammals
- C12N9/6424—Serine endopeptidases (3.4.21)
Definitions
- the invention relates to a storage-stable lysis buffer mixture for the extraction of nucleic acids from biological, preferably diagnostic samples. It is preferably associated with an extraction control.
- Areas of application include molecular biology diagnostics, research, medical practice, gene-based analysis of biotechnology, agricultural and food products as well as criminalistics.
- a large number of conventional lysis buffers for the extraction of nucleic acids contain salts as chaotropic ion mixtures.
- the associated methods are based on a method developed by Vogelstein and Gillespie (Proc Natl Acad., USA, 1979, 76, 615-619) and described for the first time for the preparative and analytical purification of DNA fragments from agarose gels.
- the method combines the dissolution of the agarose containing the DNA band to be isolated in a saturated solution of a chaotropic salt (NaJ) with a binding of the DNA to glass particles.
- NaJ chaotropic salt
- lysis buffers of this type also denature enzymes which are necessary for the lysis of a biological sample in a short time. Therefore, in conventional systems, a separate addition of the individual components must be made.
- the lysis buffer is thus assembled only during the lysis procedure and in a certain sequence, the individual components are added in succession.
- the extraction and detection sequence can be checked by two different controls: the extraction control to determine the quality of the extraction and a reaction control to determine the quality of the detection method.
- carrier nucleic acids In the case of extraction of the lowest copy numbers of nucleic acids, so-called carrier nucleic acids are frequently used, since small amounts of nucleic acids are unstable in aqueous solution. These carrier nucleic acids must not have any homology to the extracted nucleic acids and must normally be added separately in nucleic acid extraction systems. Typical carrier nucleic acids are salmon sperm DNA, herring sperm DNA, yeast t RNA and polyadenyl RNA.
- the carrier nucleic acid is usually added to the sample during the extraction. This requires an additional pipetting step.
- the extraction control is also added to the sample during extraction in a known amount. This requires a further additional pipetting step. This results in difficulties to store the control, so far as it consists of pure nucleic acids, stable in liquid form. Low concentration nucleic acids are prone to degradation.
- the extraction control is measured after the reaction by a detection method, based on the loss of control nucleic acid, the recovery rate can be determined.
- Detection method is added separately in the detection reaction, this in turn requires an additional pipetting step or approach mix for the
- Detection reaction itself contains a so-called amplification control.
- WO 0034463 Another approach is shown in WO 0034463.
- a lysis buffer based on non-chaotropic salts and suitable detergents.
- Such mixtures are, as WO 0034463 teaches, also storage-stable as solid formulations for some time.
- WO 03040386 describes the possibility of using such a system for standardized nucleic acid extraction and for standardized nucleic acid detection.
- the document WO 03040386 describes reaction spaces which have the potential for efficient and rapid processes. However, the production of such products is very complex and bound to the later reaction space.
- the preparation of the reaction spaces in the process WO 03040386 is carried out by freeze-drying.
- reaction chambers are combined with the various components of the
- the carrier nucleic acids are transferred via the lysis vessel into the reaction space for the
- Lysis buffer introduced, often together with the extraction control. This is done via so-called coating process (coating process). In this case, a solution of the nucleic acid mixture is dried on the wall of the reaction vessel. In order to complete the reaction mix for the lysis procedure, a so-called lysis buffer mix is now required.
- the lysis buffer mixture contains non-chaotropic salts and optionally
- the extraction control is also used as reaction control for the detection method.
- the sample can be added in liquid form, no further pipetting steps are necessary and an increase in the volume of liquid samples is possible.
- the production of the above system is very complex, which makes the system more expensive.
- the system only has a shelf life of six months, while the coated containers must be stored at - 20 0 C, the coated filled with Lysepuffermixtur vessels may then six months at - 20 0 C
- the object underlying the invention is therefore to provide an improved nucleic acid extraction system, which is inexpensive, stable and easy to use, while satisfying the requirements of a modern nucleic acid extraction system by u.a. Contains extraction controls.
- This object is achieved according to claims 1, 21, 23 and 24, the dependent claims are preferred variants.
- the subject matter of the present invention is a mixture, which can be prepared in liquid form, including all components, for a lysis buffer (lysis buffer mixture) for extracting nucleic acids from biological, preferably diagnostic, samples. It contains not only non-chaotropic salts for adjusting the binding conditions for the solid phase separation, detergents for lysing the biological sample, a defined amount of at least one nucleic acid as extraction control, optionally for the preparation. of low concentrations of nucleic acid so-called carrier nucleic acids, if necessary, enzymes for lysing the biological sample and optionally further additives.
- the lysis buffer mixture is used via solid phase separation for lysing the samples. It is brought into solid form via freeze-drying, thereby forming a storage-stable formulation.
- the gist of the invention is that the special detergent selection in combination with antichaotropic salts stabilizes the enzymes and nucleic acids used. It is surprising that the particular detergent selection in the lysis buffer mixture allows all components to be added together in liquid form. This leads to a mutual stabilization of the components and thereby to an increased shelf life of the freeze-dried lysis buffer mixture (at least twice as high as conventional systems) and secondly that the preparation of the corresponding mixture is independent of the reaction space.
- the lysis buffer mixture can be carried out in partial steps (eg freeze-drying) in the reaction space, but also just as completely independently thereof (eg tablet form). Thus, the later reaction can be carried out in any conceivable reaction vessel.
- Non-chaotropic salts used according to the invention are monovalent, divalent or multivalent cations or mixtures of these cations.
- these cations are ammonium ions, sodium ions, potassium ions, magnesium ions, calcium ions, zinc ions or manganese ions.
- the lysis buffer mixture is obtained by preparing two stock solutions which are mixed together, filled into reaction vessels and freeze-dried.
- the stock solution one preferably contains a mixture of one or more non-chaotropic salts, optionally tris, at least one detergent, at least one lytic enzyme.
- the stock solution two preferably contains the nucleic acid for extraction control, optionally carrier nucleic acids, sugar and TrisHCl.
- the amount of non-chaotropic salts used depends on the particular salt or mixtures.
- the salts are used to prepare the lysis buffer mixture in concentrations of 5 mmol / l - 3 mol / l.
- ammonium chloride is used as a salt, it is used in a concentration between 1 and 2 mol / l, preferably at a concentration of 1.5 mol / l.
- the pH of the lysis buffer mixture is adjusted between 5 and 9, optimally a pH of 8.
- the lysis buffer mixture according to the invention contains cetyltrimethylammonium bromide (CTAB), Tween 20, Triton X-100 and / or sodium dodecylsulfate (SDS), preferably CTAB.
- CTAB cetyltrimethylammonium bromide
- Tween 20 Triton X-100
- SDS sodium dodecylsulfate
- the concentration of detergents used for the lysis buffer mixture is dependent on the detergent and is in the range of 0.1-5%.
- CTAB is used at a concentration of 2%.
- the lysis buffer mixture contains nucleic acids for the lysis buffer mixture.
- Extraction control which are synthetic or molecular biology-produced or native nucleic acids.
- the nucleic acids can be DNA or RNA.
- the extraction control nucleic acid is used to prepare the lysis buffer mixture depending on the later detection reaction, preferably in amounts of up to 10 10 copies / ml.
- the lytic enzymes contained in the lysis buffer mixture are preferably
- proteinase K and lysozyme are used.
- nucleic acids used as carrier nucleic acids are synthetic nucleic acids or isolates of biological materials. These may preferably be poly A RNA, tRNA, salmon sperm DNA or herring sperm DNA. Others are also possible.
- the carrier nucleic acids are used for the preparation of the lysis buffer mixture in
- the lysis buffer mixture optionally contains Tris, TrisHCl and also
- sugar wherein the sugar is preferably trehalose.
- the lysis buffer mixture according to the invention is present as a solid storage-stable formulation in ready-to-use reaction vessels ready for lysing nucleic acid-containing samples.
- reaction vessels are in the specific case of the embodiments 2 ml microcentrifuge tubes, with any other type of reaction vessel is conceivable for other applications such.
- the invention further relates to a method for isolating nucleic acids from any complex starting materials using the lysis buffer mixture according to the invention.
- the method is used to isolate DNA or RNA or DNA in combination with RNA from microorganisms.
- the invention likewise relates to a reaction kit for the extraction of nucleic acids.
- the reaction kit contains vessels for sample lysis containing the lysis buffer mixture described here, optionally an alcohol-based binding buffer, optionally one or more wash buffers known per se for purifying nucleic acid, and a known elution buffer.
- the nucleic acid is purified via solid phase adsorption on silica surfaces or related surfaces.
- the kit contains spin filters, or 96-well filter plates, or magnetic beads with the corresponding surfaces.
- the reaction kit is suitable for the extraction of viral nucleic acids from diagnostic samples.
- reaction kit in another embodiment is suitable for the extraction of bacterial nucleic acids from diagnostic samples.
- the kit contains a 2 ml lysis vessel with the mixture, an isopropanol-based binding buffer, two different wash buffers known per se, an elution buffer known per se, and spin filters as the adsorption phase.
- This kit is particularly suitable for the isolation of viral nucleic acids from diagnostic samples.
- the invention has the advantage that the preparation of the lysis buffer formulation is not dependent on the reaction space.
- the mixture can therefore be produced both in the reaction space and independently.
- the extraction control nucleic acids are stabilized in a stabilizing solution (stock solution two) which is added to the lysis buffer mixture (stock solution one).
- the resulting lysis buffer mixture is used to fill vessels. This liquid combination of all components and, if necessary, an aliquoting, enables an automated production process of the formulation.
- the contents of the vessel are freeze-dried.
- the extraction control is also used in the following as a reaction control for the detection method.
- the lysis buffer mixture can be prepared in tablet form. These tablets are then added later to the sample to be lysed.
- the components After freeze-drying, the components remain stable and protect each other from any degradation that may occur. Due to their hygroscopicity, the salt components absorb to a certain extent liquid from the atmosphere, thus protecting the enzymes and the nucleic acids. Of course, it is not just the individual constituents of the mixture that cause the stabilization, but also the matched quantities of the corresponding ingredients. For the quantities, it has to be considered that the corresponding mixture must be suitable as a component in a solid-phase separation on nucleic acids.
- the lysis buffer mixture according to the invention has a shelf life of at least one year at room temperature.
- the shelf life of the lysis buffer mixtures previously described in the prior art is a maximum of six months, partly with cooling.
- lysis using the invention proceeds quite simply as follows: The sample is placed in a pre-filled vessel with the ready-mix and the volume is adjusted with water, the lysis procedure proceeds.
- WO 03040386 a lysis buffer based on non-chaotropic salts and suitable detergents.
- the production here is no longer dependent on the reaction space.
- the invention enables both automated production of the reaction unit for the lysis step at the manufacturer, as well as a simplification of automated extraction at the user by eliminating multiple liquid handling steps with small volumes (addition of enzymes, carrier nucleic acids and controls).
- An additional advantage is the increase in the possible sample volume, a liquid mixture already contains water, liquid diagnostic samples (body fluids and excreta) can be added in a larger volume. This increases the amount of isolated nucleic acids and thus the sensitivity of corresponding pathogen detections.
- this lysis buffer mixture meets the modern demands of a concentrated lysis buffer.
- the preparation of the lysis buffer mixture described here is simple, since a stabilized lysis buffer mixture is used in which all components are present together in aqueous solution.
- the sample can be added to the lysis buffer mixture in one step.
- a preparation of the lysis buffer mixture in tablet form is also possible.
- the lysis buffer mixture can likewise be added directly to the sample as a tablet.
- the lysis buffer mixture has a shelf life of at least 12 months at room temperature. The storage of this mixture is therefore possible without cooling.
- the sample can be added in liquid form, no further pipetting steps are necessary, thereby reducing the risk of contamination for the sample by cross-contamination and also for the user on the basis of infectious samples.
- a solution I of 1.5 M ammonium chloride, 10 mM Tris pH 8, 2% CTAB, 0.5 mg proteinase K / ml and 0.5 mg lysozyme / ml is prepared.
- Solution I and II are mixed in a ratio of 40: 1
- Embodiment 2 The solution is frozen in 400 ⁇ l portions in sealable 2 ml reaction vessels. Then these containers are freeze-dried with content, they are called in the text text Extraction Tubes Bacteria. Embodiment 2
- Fragment plasmid pCONT 10 8 copies / ml control RNA fragment, 1% trehalose and 50 mM TrisCl, pH 8.
- Solution I and II are mixed in a ratio of 40: 1
- the solution is frozen in 400 ⁇ l portions in sealable 2 ml reaction vessels. Then these containers are freeze-dried with content, they are called in the text Extraction Tube Virus.
- Serum samples 200 ⁇ l
- HBV 500 copies per preparation or in estimated titer number (influenza A virus) are used for the extraction.
- the spin filter is washed twice with a wash buffer consisting of 10 mM Tris pH8, 70% ethanol.
- the spin filter is centrifuged dry for five minutes. It is eluted with 100 .mu.l RNase / Dnase free water while preincubated for three minutes and then centrifuged.
- the viruses and the extraction controls are detected by quantitative PCR systems or by quantitative reverse transcriptase PCR systems.
- RNA Control Detection Assay RNA Control Detection Assay
- Serum samples 200 ⁇ l
- HBV 500 copies per preparation or in estimated titer number (influenza A virus) are used for the extraction.
- viruses and the extraction controls are detected by quantitative PCR systems or quantitative reverse transcriptase PCR systems.
- quantitative PCR systems For influenza A and HBV detection systems from Gongen are used
- RNA Control Detection Assay RNA Control Detection Assay
- the magnetic particles are mixed by a magnetic separator, KingFisher mL, from Thermo. Then the magnetic particles are washed in two successive wells with the wash buffer consisting of 10 mM Tris pH8, 70% ethanol. The magnetic particles are then dried for 10 minutes at room temperature to remove the alcohol. It is eluted with 100 .mu.l RNase / Dnase free water in another well, mixed for three minutes and then the magnetic particles are aseparated. The bacteria and the extraction control are detected by quantitative PCR systems.
Abstract
Description
Claims
Priority Applications (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
DE112009001513.1T DE112009001513B4 (de) | 2008-04-22 | 2009-04-20 | Stabile Lysepuffermixtur zur Extraktion von Nukleinsäuren |
US12/988,906 US20110092687A1 (en) | 2008-04-22 | 2009-04-20 | Stable lysis buffer mixture for extracting nucleic acids |
US14/171,157 US9593325B2 (en) | 2008-04-22 | 2014-02-03 | Stable lysis buffer mixture for extracting nucleic acids |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
DE102008020258A DE102008020258A1 (de) | 2008-04-22 | 2008-04-22 | Stabile Lysepuffermixtur zur Extraktion von Nukleinsäuren |
DE102008020258.4 | 2008-04-22 |
Related Child Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US12/988,906 A-371-Of-International US20110092687A1 (en) | 2008-04-22 | 2009-04-20 | Stable lysis buffer mixture for extracting nucleic acids |
US14/171,157 Continuation US9593325B2 (en) | 2008-04-22 | 2014-02-03 | Stable lysis buffer mixture for extracting nucleic acids |
Publications (3)
Publication Number | Publication Date |
---|---|
WO2009129797A2 true WO2009129797A2 (de) | 2009-10-29 |
WO2009129797A3 WO2009129797A3 (de) | 2009-12-30 |
WO2009129797A4 WO2009129797A4 (de) | 2010-03-04 |
Family
ID=41111646
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
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PCT/DE2009/000549 WO2009129797A2 (de) | 2008-04-22 | 2009-04-20 | Stabile lysepuffermixtur zur extraktion von nukleinsäuren |
Country Status (3)
Country | Link |
---|---|
US (2) | US20110092687A1 (de) |
DE (2) | DE102008020258A1 (de) |
WO (1) | WO2009129797A2 (de) |
Families Citing this family (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE102011054474B4 (de) | 2011-07-20 | 2014-02-13 | Stratec Biomedical Ag | System zur Stabilisierung, Aufbewahrung und Lagerung einer Nukleinsäure |
CN105074003A (zh) | 2012-09-13 | 2015-11-18 | 哈佛大学校长及研究员协会 | 实体组织的单细胞中的多元分析测量的方法 |
EP3739062B1 (de) | 2014-10-20 | 2023-08-16 | Gen-Probe Incorporated | Lyselösung roter blutkörperchen |
EP3448998B1 (de) | 2016-04-27 | 2020-06-03 | Gen-Probe Incorporated | Reagens für blutzelllyse |
WO2018063772A1 (en) * | 2016-09-27 | 2018-04-05 | Abbott Molecular Inc. | Maximizing dna yield of blood specimens collected in rapid clot tubes |
US11647993B2 (en) * | 2017-12-22 | 2023-05-16 | Research Triangle Institute | Oral fluid collector |
CN111778236A (zh) * | 2020-06-23 | 2020-10-16 | 辽宁省海洋水产科学研究院 | 基于3d打印特形功能体的贝类基因组dna提取方法、试剂盒及其应用 |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
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WO2003040386A2 (de) * | 2001-11-02 | 2003-05-15 | InViTek Gesellschaft für Biotechnik & Biodesign mbH | Reaktionsräume enthaltend komplexe lagerstabile reagenzienformulierungen und testkit zum nachweis und zur isolierung pathogener mikrobieller nukleinsäuren |
EP1462520A1 (de) * | 2003-03-28 | 2004-09-29 | Eppendorf 5 Prime, Inc. | Verfahren zur Isolierung von DNA |
EP1529840A1 (de) * | 2003-11-04 | 2005-05-11 | Qiagen GmbH | Ein schnelles und preiswertes Verfahren zur Gewinnung von Nukleinsäuren |
DE102006019650A1 (de) * | 2006-04-25 | 2007-10-31 | InViTek Gesellschaft für Biotechnik & Biodesign mbH | Formulierungen und Verfahren zur Isolierung von Nukleinsäuren aus beliebigen komplexen Ausgangsmaterialien und nachfolgende komplexe Genanalytik |
Family Cites Families (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5234809A (en) | 1989-03-23 | 1993-08-10 | Akzo N.V. | Process for isolating nucleic acid |
DE59912604D1 (de) * | 1998-02-04 | 2005-11-03 | Merck Patent Gmbh | Verfahren zur isolierung und aufreinigung von nucleinsäuren |
DE19840531C2 (de) | 1998-08-28 | 2003-05-15 | Roboscreen Ges Fuer Molekulare | Mit Nukleinsäuren beschichtete Reaktionsräume, Verfahren zu ihrer Herstellung und ihre Verwendung |
DE19856064C2 (de) | 1998-12-04 | 2000-11-30 | Invitek Gmbh | Universelles Verfahren zur Isolierung von DNA aus beliebigen Ausgangsmaterialien |
WO2001013086A2 (en) * | 1999-08-13 | 2001-02-22 | Brandeis University | Detection of nucleic acids |
US8852862B2 (en) * | 2004-05-03 | 2014-10-07 | Handylab, Inc. | Method for processing polynucleotide-containing samples |
US20060223071A1 (en) * | 2005-04-01 | 2006-10-05 | Wisniewski Michele E | Methods, compositions, and kits for detecting nucleic acids in a single vessel |
US8187557B2 (en) * | 2006-07-13 | 2012-05-29 | Cepheid | Reagent reservoir system for analytical instruments |
-
2008
- 2008-04-22 DE DE102008020258A patent/DE102008020258A1/de not_active Withdrawn
-
2009
- 2009-04-20 US US12/988,906 patent/US20110092687A1/en not_active Abandoned
- 2009-04-20 WO PCT/DE2009/000549 patent/WO2009129797A2/de active Application Filing
- 2009-04-20 DE DE112009001513.1T patent/DE112009001513B4/de active Active
-
2014
- 2014-02-03 US US14/171,157 patent/US9593325B2/en active Active
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2003040386A2 (de) * | 2001-11-02 | 2003-05-15 | InViTek Gesellschaft für Biotechnik & Biodesign mbH | Reaktionsräume enthaltend komplexe lagerstabile reagenzienformulierungen und testkit zum nachweis und zur isolierung pathogener mikrobieller nukleinsäuren |
EP1462520A1 (de) * | 2003-03-28 | 2004-09-29 | Eppendorf 5 Prime, Inc. | Verfahren zur Isolierung von DNA |
EP1529840A1 (de) * | 2003-11-04 | 2005-05-11 | Qiagen GmbH | Ein schnelles und preiswertes Verfahren zur Gewinnung von Nukleinsäuren |
DE102006019650A1 (de) * | 2006-04-25 | 2007-10-31 | InViTek Gesellschaft für Biotechnik & Biodesign mbH | Formulierungen und Verfahren zur Isolierung von Nukleinsäuren aus beliebigen komplexen Ausgangsmaterialien und nachfolgende komplexe Genanalytik |
Also Published As
Publication number | Publication date |
---|---|
US9593325B2 (en) | 2017-03-14 |
WO2009129797A4 (de) | 2010-03-04 |
US20140154776A1 (en) | 2014-06-05 |
DE112009001513A5 (de) | 2011-04-07 |
DE112009001513B4 (de) | 2016-03-17 |
WO2009129797A3 (de) | 2009-12-30 |
US20110092687A1 (en) | 2011-04-21 |
DE102008020258A1 (de) | 2009-10-29 |
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