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WO1994029005A1 - Dispositif d'electro-elution a puits d'agarose - Google Patents

Dispositif d'electro-elution a puits d'agarose Download PDF

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Publication number
WO1994029005A1
WO1994029005A1 PCT/KR1994/000063 KR9400063W WO9429005A1 WO 1994029005 A1 WO1994029005 A1 WO 1994029005A1 KR 9400063 W KR9400063 W KR 9400063W WO 9429005 A1 WO9429005 A1 WO 9429005A1
Authority
WO
WIPO (PCT)
Prior art keywords
agarose
nucleic acids
trap
gel
plugged
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/KR1994/000063
Other languages
English (en)
Inventor
Yeon Bo Chung
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.)
Individual
Original Assignee
Individual
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 Individual filed Critical Individual
Publication of WO1994029005A1 publication Critical patent/WO1994029005A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/416—Systems
    • G01N27/447—Systems using electrophoresis
    • G01N27/44704—Details; Accessories
    • G01N27/44717—Arrangements for investigating the separated zones, e.g. localising zones
    • G01N27/4473—Arrangements for investigating the separated zones, e.g. localising zones by electric means
    • 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
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • 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
    • 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
    • B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
    • B01L2400/0415—Moving fluids with specific forces or mechanical means specific forces electrical forces, e.g. electrokinetic
    • B01L2400/0421—Moving fluids with specific forces or mechanical means specific forces electrical forces, e.g. electrokinetic electrophoretic flow

Definitions

  • Nucleic acids are usually separated by electrophoresis on agarose or acrylamide gels. Purification of separated nucleic acid from the gel matrix is an obligatory and critical step in most of the molecular biological research. Diverse methods have been developed to recover the fractionated nucleic acids (1, 2). A common approach is the electroelution of nucleic acids out of the gel slice into the solution. Samples can be eluted into a dialysis bag, to a high-salt layer maintained in a V-channel using an apparatus commercially available (Electroeluter, IBI, USA) or to a DEAE-ion- exchange paper embedded in the trough just ahead of the target band. The bound DNA is later recovered by alcohol-precipitation. Another approach is the removal of gel matrix. Specially prepared agarose with low-gelling temperature (SeaPlaque, FMC,
  • agarose Ordinary agarose could also be solubilized at room temperature in the presence of potassium idodide and recovered by adsorption to glass powder (GeneClean, BIOlOl, USA). Enzymatic digestion of agarose is also possible ( ⁇ -Agarase, New
  • a model of agarose-plugged electroeluter and details of the eluter trap The agarose-plugged electroeluter is composed of three major parts; upper buffer reservoir, trap part, and lower reservoir. A platinum wire (1, 2) is placed on each reservoir. The overall dimension is 12 cm (width) x 8 cm (depth) x 15 cm (height) and is made of acryl.
  • the trap part(3) is permanently attached to the upper reservoir(4). Half of the trap is protruding the bottom of the upper reservoir to be immersed in the buffer of the lower reservoir(5).
  • the two reservoirs are connected solely by the narrow channels(6) in the trap part(3).
  • the triangle-shaped upper part of the trap is the gel-receptacle(7). It leads to the tiered narrow channels (8, 9) below.
  • the lower oriface(l ⁇ ) is plugged with 1% agarose at the beginning, and nucleic-acid-trapping material fills the cylinder. We recommend 9 M ammonium acetate or DEAE-cellulose resin for this trap. Then the upper oriface is plugged with melt agarose simultaneously with the placement of the sample gel slice to get embedded in the plug.
  • the tiered structure of the channel is to prevent the slippage of the agarose plug.
  • the slope(14) of the bottom surface of the trap part is to prevent the accumulation of air bubbles.
  • the apparatus can be build based on the above design and distributred commercially to scientists and engineers involved in life science.
  • the apparatus should be manufactured in a factory with automatic processing.
  • the apparatus is an analytical device and intended for use in indivisual laboratories. It is not for mass purification or isolation of biological material.

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  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Analytical Chemistry (AREA)
  • Organic Chemistry (AREA)
  • Genetics & Genomics (AREA)
  • General Health & Medical Sciences (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Molecular Biology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biochemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • General Engineering & Computer Science (AREA)
  • Physics & Mathematics (AREA)
  • Biophysics (AREA)
  • Microbiology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Immunology (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Plant Pathology (AREA)
  • Electrochemistry (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Hematology (AREA)
  • Clinical Laboratory Science (AREA)
  • Saccharide Compounds (AREA)

Abstract

L'isolation d'acides nucléiques est une étape fondamentale et critique de la plupart des études biologiques moléculaires. Les acides nucléiques sont généralement fractionnés par taille sur une matrice de gel par électrophorèse et ils doivent être purifiés de cette matrice en vue de leur manipulation et de leur analyse ultérieures. L'électro-élution est un procédé permettant d'extraire, par circulation d'un courant électrique, des acides nucléiques de la matrice de gel pour les faire passer dans une solution. L'invention décrit un nouveau modèle de dispositif d'électro-élution. Un cylindre rempli d'une substance susceptible de maintenir stables des acides nucléiques est pourvu à chaque extrémité de puits d'agarose. Des lamelles échantillons de gel sont disposées de manière à permettre une fusion avec le puits d'agarose supérieur côté cathode. L'acide nucléique quitte la lamelle de gel pour pénétrer dans le cylindre où il est maintenu par la substance-piège. Lorsque l'élution est terminée, le puits supérieur est ôté et la substance-piège est recueillie conjointement avec l'acide nucléique. Ce dernier est ensuite récupéré par précipitation avec de l'alcool.
PCT/KR1994/000063 1993-06-09 1994-06-03 Dispositif d'electro-elution a puits d'agarose Ceased WO1994029005A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
KR1993/10214 1993-06-09
KR2019930010214U KR950001267U (ko) 1993-06-09 1993-06-09 전기핵산유출기

Publications (1)

Publication Number Publication Date
WO1994029005A1 true WO1994029005A1 (fr) 1994-12-22

Family

ID=60917523

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR1994/000063 Ceased WO1994029005A1 (fr) 1993-06-09 1994-06-03 Dispositif d'electro-elution a puits d'agarose

Country Status (2)

Country Link
KR (1) KR950001267U (fr)
WO (1) WO1994029005A1 (fr)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6942775B1 (en) * 2002-05-24 2005-09-13 Owl Separation Systems, Inc. Vertical electrophoresis system
US7037419B2 (en) * 2001-09-14 2006-05-02 Peter James Concentration of protein and/or peptides samples
WO2007015625A1 (fr) * 2005-08-01 2007-02-08 Jae Gyeong Jeong Système de récupération d’adn et arn ou de fragments de protéines avec un gel d’agarose ou un gel polyacrylamide
WO2012171329A1 (fr) * 2011-06-15 2012-12-20 Du Quan Procédé de séparation d'un acide nucléique et ses applications

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4391688A (en) * 1981-06-02 1983-07-05 Institut Armand-Frappier Electrophoresis system for multiple agarose slab gels
DE3715856A1 (de) * 1987-05-12 1988-12-01 Biometra Biomedizinische Analy Vorrichtung zum konzentrieren von in einer fluessigkeit befindlichen, elektrisch geladenen - insbesondere aus einem gel eluierten - makromolekuelen
EP0459241A1 (fr) * 1990-05-29 1991-12-04 Waters Investments Limited Procédé et dispositif pour l'exécution d'électrophorèse capillaire

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4391688A (en) * 1981-06-02 1983-07-05 Institut Armand-Frappier Electrophoresis system for multiple agarose slab gels
DE3715856A1 (de) * 1987-05-12 1988-12-01 Biometra Biomedizinische Analy Vorrichtung zum konzentrieren von in einer fluessigkeit befindlichen, elektrisch geladenen - insbesondere aus einem gel eluierten - makromolekuelen
EP0459241A1 (fr) * 1990-05-29 1991-12-04 Waters Investments Limited Procédé et dispositif pour l'exécution d'électrophorèse capillaire

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7037419B2 (en) * 2001-09-14 2006-05-02 Peter James Concentration of protein and/or peptides samples
US6942775B1 (en) * 2002-05-24 2005-09-13 Owl Separation Systems, Inc. Vertical electrophoresis system
WO2007015625A1 (fr) * 2005-08-01 2007-02-08 Jae Gyeong Jeong Système de récupération d’adn et arn ou de fragments de protéines avec un gel d’agarose ou un gel polyacrylamide
WO2012171329A1 (fr) * 2011-06-15 2012-12-20 Du Quan Procédé de séparation d'un acide nucléique et ses applications

Also Published As

Publication number Publication date
KR950001267U (ko) 1995-01-04

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