WO2016169677A1 - Verfahren und testkit zur schnellen isolierung von nukleinsäuren mittels rauer oberflächen - Google Patents

Verfahren und testkit zur schnellen isolierung von nukleinsäuren mittels rauer oberflächen Download PDF

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Publication number
WO2016169677A1
WO2016169677A1 PCT/EP2016/054178 EP2016054178W WO2016169677A1 WO 2016169677 A1 WO2016169677 A1 WO 2016169677A1 EP 2016054178 W EP2016054178 W EP 2016054178W WO 2016169677 A1 WO2016169677 A1 WO 2016169677A1
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WO
WIPO (PCT)
Prior art keywords
nucleic acids
plastic
solid phase
nucleic acid
rough
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.)
Ceased
Application number
PCT/EP2016/054178
Other languages
German (de)
English (en)
French (fr)
Inventor
Timo Hillebrand
Thorsten Stroh
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.)
AJ Innuscreen GmbH
Original Assignee
AJ Innuscreen 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
Priority claimed from DE102015211393.0A external-priority patent/DE102015211393A1/de
Priority claimed from DE102015211394.9A external-priority patent/DE102015211394B4/de
Priority to CA2983619A priority Critical patent/CA2983619C/en
Application filed by AJ Innuscreen GmbH filed Critical AJ Innuscreen GmbH
Priority to JP2018506477A priority patent/JP6979940B2/ja
Priority to CN201680036533.2A priority patent/CN108124454A/zh
Priority to EP16711778.7A priority patent/EP3286325B1/de
Priority to US15/568,475 priority patent/US10934540B2/en
Priority to PL16711778.7T priority patent/PL3286325T3/pl
Priority to ES16711778T priority patent/ES2928055T3/es
Publication of WO2016169677A1 publication Critical patent/WO2016169677A1/de
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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Classifications

    • 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
    • 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
    • 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
    • B01L3/02—Burettes; Pipettes
    • B01L3/0275—Interchangeable or disposable dispensing tips
    • 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
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/40—Concentrating samples
    • G01N1/405—Concentrating samples by adsorption or absorption
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/10—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices
    • 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
    • B01L2200/02—Adapting objects or devices to another
    • B01L2200/023—Adapting objects or devices to another adapted for different sizes of tubes, tips or container
    • 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
    • B01L2200/0631—Purification arrangements, e.g. solid phase extraction [SPE]
    • 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
    • B01L2200/0647—Handling flowable solids, e.g. microscopic beads, cells, particles
    • 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
    • B01L2300/0848—Specific forms of parts of containers
    • B01L2300/0854—Double walls
    • 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
    • B01L2300/16—Surface properties and coatings
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10T—TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
    • Y10T436/00—Chemistry: analytical and immunological testing
    • Y10T436/25—Chemistry: analytical and immunological testing including sample preparation
    • Y10T436/25375—Liberation or purification of sample or separation of material from a sample [e.g., filtering, centrifuging, etc.]
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10T—TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
    • Y10T436/00—Chemistry: analytical and immunological testing
    • Y10T436/25—Chemistry: analytical and immunological testing including sample preparation
    • Y10T436/25375—Liberation or purification of sample or separation of material from a sample [e.g., filtering, centrifuging, etc.]
    • Y10T436/255—Liberation or purification of sample or separation of material from a sample [e.g., filtering, centrifuging, etc.] including use of a solid sorbent, semipermeable membrane, or liquid extraction

Definitions

  • the invention was based on the following observation.
  • a sample containing nucleic acid with extraction reagents from commercially available kits e.g.
  • the subsequent steps of washing can be carried out with known alcohol-containing washing buffers or even with an alcohol, as well as known for the elution low-salt buffer or water can be used.
  • the essence of the invention is thus that free or liberated by lysis nucleic acids are in an aqueous environment whose polarity is adjusted by means of organic substances so that the solubility of the nucleic acid is reduced and subsequently this aqueous environment with a rough surface is brought into contact so that then precipitates the nucleic acid on the rough surface and subsequently the precipitated DNA is peeled off again from the rough surface and is available.
  • the precipitated on the rough surface nucleic acid can also be washed and replaced after washing steps.
  • the present invention thus makes it possible in the most ideal way, the implementation of the objectives already listed.
  • All that is required for the extraction is a material whose surface is roughened and an aqueous environment in which the nucleic acid to be isolated is, the conditions of which by means of an organic substance are adjusted such that the solubility of the nucleic acid is reduced to that of the rough Surface precipitated.
  • This precipitation on the rough surface is done by Contacting the rough surface with the sample or contacting the sample with the rough surface.
  • classical kit reagents eg lysis buffers
  • the necessary conditions for the precipitation of the nucleic acid to the rough surface are adjusted by the addition of an organic substance.
  • the extraction protocol works according to the known scheme lysing-binding-washing-eluting. Alternatively, you can
  • Cavity 2 800 ⁇ Washing Solution LS (from the extraction kit)
  • Cavity 2 800 ⁇ Washing Solution LS (from the extraction kit)
  • FIG. 2 shows the detection of the DNA on an agarose gel. 4 ⁇ nucleic acid from the total eluate of 200 ⁇ were used per lane. The tracks show: 1. DNA ladder (lb ladder)

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  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Genetics & Genomics (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Analytical Chemistry (AREA)
  • Biomedical Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Physics & Mathematics (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Biophysics (AREA)
  • Microbiology (AREA)
  • Molecular Biology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Immunology (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Plant Pathology (AREA)
  • Clinical Laboratory Science (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Treatment Of Liquids With Adsorbents In General (AREA)
  • Solid-Sorbent Or Filter-Aiding Compositions (AREA)
  • Sampling And Sample Adjustment (AREA)
  • Saccharide Compounds (AREA)
PCT/EP2016/054178 2015-04-23 2016-02-26 Verfahren und testkit zur schnellen isolierung von nukleinsäuren mittels rauer oberflächen Ceased WO2016169677A1 (de)

Priority Applications (7)

Application Number Priority Date Filing Date Title
ES16711778T ES2928055T3 (es) 2015-04-23 2016-02-26 Procedimiento y kit de test para el aislamiento rápido de ácidos nucleicos por medio de superficies rugosas
PL16711778.7T PL3286325T3 (pl) 2015-04-23 2016-02-26 Sposób i zestaw testowy do izolowania kwasów nukleinowych za pomocą szorstkich powierzchni
CA2983619A CA2983619C (en) 2015-04-23 2016-02-26 Method and test kit for rapid isolation of nucleic acids using rough surfaces
JP2018506477A JP6979940B2 (ja) 2015-04-23 2016-02-26 粗さのある表面による核酸の迅速な単離のための方法および試験キット
CN201680036533.2A CN108124454A (zh) 2015-04-23 2016-02-26 借助粗糙表面快速分离核酸的方法和试剂盒
EP16711778.7A EP3286325B1 (de) 2015-04-23 2016-02-26 Verfahren und testkit zur schnellen isolierung von nukleinsäuren mittels rauer oberflächen
US15/568,475 US10934540B2 (en) 2015-04-23 2016-02-26 Method and test kit for rapid isolation of nucleic acids using rough surfaces

Applications Claiming Priority (6)

Application Number Priority Date Filing Date Title
DE102015207481 2015-04-23
DE102015207481.1 2015-04-23
DE102015211393.0A DE102015211393A1 (de) 2015-06-19 2015-06-19 Vorrichtung und Verfahren zur automatisierten Extraktion von Nukleinsäuren
DE102015211394.9A DE102015211394B4 (de) 2015-06-19 2015-06-19 Vorrichtung und Verfahren zur Extraktion von Nukleinsäuren
DE102015211393.0 2015-06-19
DE102015211394.0 2015-06-19

Publications (1)

Publication Number Publication Date
WO2016169677A1 true WO2016169677A1 (de) 2016-10-27

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Application Number Title Priority Date Filing Date
PCT/EP2016/054180 Ceased WO2016169679A1 (de) 2015-04-23 2016-02-26 Vorrichtung und verfahren zur automatisierten extraktion von nukleinsäuren
PCT/EP2016/054178 Ceased WO2016169677A1 (de) 2015-04-23 2016-02-26 Verfahren und testkit zur schnellen isolierung von nukleinsäuren mittels rauer oberflächen
PCT/EP2016/054179 Ceased WO2016169678A1 (de) 2015-04-23 2016-02-26 Vorrichtung und verfahren zur extraktion von nukleinsäuren

Family Applications Before (1)

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PCT/EP2016/054180 Ceased WO2016169679A1 (de) 2015-04-23 2016-02-26 Vorrichtung und verfahren zur automatisierten extraktion von nukleinsäuren

Family Applications After (1)

Application Number Title Priority Date Filing Date
PCT/EP2016/054179 Ceased WO2016169678A1 (de) 2015-04-23 2016-02-26 Vorrichtung und verfahren zur extraktion von nukleinsäuren

Country Status (8)

Country Link
US (3) US10851368B2 (en:Method)
EP (3) EP3286313A1 (en:Method)
JP (4) JP6797185B2 (en:Method)
CN (3) CN108076644B (en:Method)
CA (3) CA2983619C (en:Method)
ES (2) ES2928055T3 (en:Method)
PL (2) PL3286312T3 (en:Method)
WO (3) WO2016169679A1 (en:Method)

Cited By (5)

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WO2021013534A1 (de) 2019-07-07 2021-01-28 Aj Innuscreen Gmbh Verfahren und testkit zur bisulfitmodifizierung von dna
DE102020135124A1 (de) 2020-12-30 2022-06-30 Aj Innuscreen Gmbh Verfahren und System zur schnellen Isolierung von Nukleinsäuren direkt aus Vollblutproben
DE102021104319A1 (de) 2021-02-23 2022-08-25 Aj Innuscreen Gmbh Verfahren zur simultanen Isolierung von Nukleinsäuren aus sedimentierbaren und nicht sedimentierbaren Biomolekülen in Wasserproben
DE102021130283A1 (de) 2021-11-19 2023-05-25 Ist Innuscreen Gmbh Verfahren und testkit zur preiswerten und ressourcensparenden extraktion von nukleinsäuren
WO2025141189A1 (de) * 2023-12-29 2025-07-03 Ist Innuscreen Gmbh Verfahren, mittel und kit zur extraktion hochmolekularer dna

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EP3286313A1 (de) 2015-04-23 2018-02-28 AJ Innuscreen GmbH Vorrichtung und verfahren zur automatisierten extraktion von nukleinsäuren
DE102017204267B4 (de) * 2017-03-14 2021-05-27 Aj Innuscreen Gmbh Verfahren zur anreicherung von zellen aus einer probe und der nachfolgenden nukleinsäureisolierung aus diesen zellen
US12264311B2 (en) 2019-01-25 2025-04-01 Sekisui Chemical Co., Ltd. Method for isolating nucleic acid, nucleic acid isolation kit, and inspection chip
DE102019201966B4 (de) * 2019-02-14 2023-07-27 Axagarius Gmbh & Co. Kg Probentransferwerkzeug
CN113546703A (zh) * 2021-07-30 2021-10-26 苏州含光微纳科技有限公司 一种离心式微流控芯片
CN113820195B (zh) * 2021-11-25 2022-03-08 成都宜乐芯生物科技有限公司 一种多通道并行前处理装置
DE102022115445B4 (de) 2022-06-21 2024-07-04 Ist Innuscreen Gmbh Verfahren und kit zur manuellen und automatisierten probenvorbereitung von long-read-sequenzierungen
CN115141719A (zh) * 2022-07-13 2022-10-04 安徽百奥秘科生物医药研究院有限公司 一种无阻碍吸入式动态快速提取核酸的方法及装置

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