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WO2002063042A1 - Apparatus for isolating nucleic acid or biological material - Google Patents

Apparatus for isolating nucleic acid or biological material Download PDF

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Publication number
WO2002063042A1
WO2002063042A1 PCT/KR2001/000153 KR0100153W WO02063042A1 WO 2002063042 A1 WO2002063042 A1 WO 2002063042A1 KR 0100153 W KR0100153 W KR 0100153W WO 02063042 A1 WO02063042 A1 WO 02063042A1
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WO
WIPO (PCT)
Prior art keywords
slider
solid material
chambers
slot
chamber
Prior art date
Application number
PCT/KR2001/000153
Other languages
French (fr)
Inventor
Gi Young Jang
Original Assignee
Bionex, Inc.
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 Bionex, Inc. filed Critical Bionex, Inc.
Priority to PCT/KR2001/000153 priority Critical patent/WO2002063042A1/en
Publication of WO2002063042A1 publication Critical patent/WO2002063042A1/en

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Classifications

    • 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
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • 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
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54366—Apparatus specially adapted for solid-phase testing
    • 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
    • B01L2200/0605—Metering of fluids
    • 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
    • B01L2400/0475—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure
    • 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
    • B01L2400/06—Valves, specific forms thereof
    • B01L2400/0633—Valves, specific forms thereof with moving parts
    • B01L2400/065—Valves, specific forms thereof with moving parts sliding valves

Definitions

  • the present invention relates to an apparatus for isolating and purifying nucleic acid, cell and various biological materials.
  • the advantages of the method using these materials is that no harmful organic solvent is involved, and physical and biochemical degrading of nucleic acid during the isolation process is minimized.
  • immobilized nucleic acid is less susceptible to digestion by nucleic acid degrading enzymes.
  • a feature of the present invention is to provide an apparatus for isolating a nucleic or biological material from low and medium number of biological samples in which intensive manual pipetting step is eliminated and the risk of viral and bacterial infections from the biological samples is reduced.
  • Another feature of the present invention is to provide an apparatus for isolating nucleic acid or biological material from the biological samples less expensive and more readily available to researchers, medical personnel and other potential users of the technology.
  • an apparatus for isolating nucleic acid or biological material from biological sample using a solid material which carries or absorbs nucleic acid or biological material comprising:
  • an apparatus for isolating nucleic acid or biological material from biological sample using a solid material which carries or absorbs nucleic acid or biological material comprising:
  • the various biological samples can be applied over isolation of nucleic acid or biological materials.
  • the various biological samples are blood, bacteria, cultured cells, polymerase chain Reaction (PCR) samples, DNA sequencing samples, DNA-containing agarose product and other biological reaction samples.
  • the examples of the solid material absorbing or binding nucleic acid or biological materials are silica, glass fiber, anion exchange resins, natural magnetic bead, modified magnetic bead, Sephadex, Sepharose and Sephacryl.
  • Some solid material such as silica, glass fibers, anion exchange resins, Sephadex, Sephacryl and Sepharose have the binding ability to nucleic acid or biological material and can be settled down somewhat rapidly under gravity.
  • the other solid material such as natural or modified magnetic bead have the binding ability to nucleic acid or biological materials depending on modification on surface and affinity to magnet. The feature of the solid material allows the collection of the solid material into the cavity of the slider.
  • FIG. 1 is a cross sectional view illustrating sequential steps of isolating a nucleic or biological materials using an apparatus according to the present invention
  • FIG. 2 is a top view of an apparatus according to the present invention in which a cassette contains eight sets of plural chambers;
  • FIG. 3 is a cross sectional view of an apparatus according to one example of the present invention.
  • FIG. 4 is a cross sectional view of an apparatus according to another example of the present invention.
  • the present invention provides an apparatus for isolating nucleic acid or biological material from various biological samples.
  • the feature in the present invention is that solid material such as silica, glass fibers, exchange resin, modified magnetic beads and etc. is used for binding or absorbing nucleic acid or biological materials to remove the use of organic solvents and the centrifugation step.
  • a slider is applied for efficient movement of solid material binding nucleic acid or biological materials.
  • FIG. 1 is a cross sectional view illustrating sequential steps of isolating a nucleic or biological materials using an apparatus according to a preferred embodiment of the present invention.
  • a cassette 100 comprises four chambers 102a,
  • the four chambers 102a, 102b, 102c and 102d are connected with one another with a slot which is fitted with a slider 105.
  • the slider 105 can be pulled back and forth according to the length of the slot, the slider 105 fits into the slot very tightly so that the spaces of the chambers 102a, 102b, 102c and 102d are separated and the liquid reactants 103a, 103b, 103c and 103d which is added into the chambers 102a, 102b, 102c and 102d are sustained respectively.
  • the slider 105 has a small cavity 106 that is capable to settle or collect a solid material at one of its end parts.
  • the depth, area and shape of the cavity can be varied depending on the kind of used solid material and the shape of the chambers 102a, 102b, 102c and 102d.
  • the cross sectional shape of the cavity 106 is the same with that of the chambers 102a, 102b, 102c and 102d.
  • the liquid reactants 103a, 103b, 103c and 103d in each chambers 102a, 102b, 102c and 102d can be various buffers such as lysis buffer, ethanol elution buffer, distilled water and etc. which is needed in the process of isolating nucleic acid or biological materials.
  • the process starts with an addition of blood into the chamber 102a that already contains lysis buffer 103a and a magnetic bead 104.
  • the whole cassette 100 is shaken, for example, back and forth, or in seesawing or rotating motion in conjunction with motioning system (not shown).
  • a magnet 107 is applied to collect the DNA-carrying magnetic bead 104 into the cavity 106 of the slider 105 as shown in FIG.
  • the slider 105 is pulled manually or by automatic pulling system driven by a motor (not shown) and the cavity 106 is located in the bottom of the chamber 102b as shown in FIG. 1(b). After that, for washing the magnetic bead 104, the whole cassette
  • the liquid reactant 103d is an elution buffer to elute the nucleic acid from magnetic bead 104 and finally the liquid reactant 103d containing nucleic acid in the last chamber 102d is collected by pipetting through an input hole lOld.
  • the input hole 101a, 101b, 101c and lOld was shown up in a simplified format in FIG. 1 , other input methods such as a self-sealing piercable diaphragm, and etc. can be used.
  • the input holes 101a, 101b, 101c and lOld can be equipped with a sliding lid to avoid flowing of the liquid reactants 103a, 103b, 103c and 103d.
  • the step of magnet 107 application to collect the magnetic bead 104 can be omitted if the other type of solid materials such as silica and resins that are easily settled down by gravity is used instead of the magnetic bead 104.
  • a cassette 201 contains eight sets of the chambers 202 for isolating nucleic acid or biological materials from eight different samples.
  • a slider 205 can be pulled according to the direction of the arrow.
  • the number of the chamber set can be varied from 1 to 20,000.
  • the number of chamber set can be increased by preferably reducing the size of each chamber or enlarging the size of the cassette.
  • the shape of each chamber can be varied within limited area of the chamber.
  • FIG. 3 is a cross sectional view of an apparatus according to one embodiment of the present invention.
  • the apparatus is explained focused on a means for pulling the slider, a means for generating magnetic force, and a means for agitating the liquid reactants and the solid material.
  • the means for pulling the slider comprises a belt 314, a hook 313 that fixes the slider to the belt 314, a pivoting point 312 to hold one side of the belt 314, and a motor 315 that drives the belt 314. Accordingly, the slider is pulled by driving the motor 315.
  • the means for generating magnetic force comprises a magnet 318, a belt 317, a pivoting point 316 to hold one side of the belt 317 and a motor 319 to drives the belt. Thereby, a magnet can be moved according to the movement of magnetic solid material.
  • the means for controlling the temperature is a heater 320 which is operated by electrical power and controlled by a censor 321.
  • FIG. 4 is a cross sectional view of an apparatus according to the other embodiment of the present invention.
  • the main difference of the apparatus in FIG. 4 from the apparatus in FIG. 3 is addition of a second slider 408 on the upper part of a cassette 400 instead of seesawing part, and a system to pull the cassette body 400 and a cassette holder 408 instead of the system to pull a slider 405 and a magnet 407.
  • the input holes are omitted.
  • the second slider 408 is a flat plate having multi number of vertical intruder 410. The purpose of the second slider 408 is to agitate the liquid reactants and the solid material in the plural chambers.
  • the second slider 408 has an advantage especially for the case that the mixing reaction is well not achieved by shaking the cassette since the liquid reactants have high surface tension.
  • the liquid reactants and the solid material are agitated by pulling the second slider 408 back and forth repeatedly.
  • a means for pulling the second slider 408 comprises a belt (not shown), a hook 413 that fixes the slider to the belt, and a motor (not shown) that drives the belt.
  • Movement of the solid material on the cavity is achieved by moving the cassette 400, the cassette holder 409, and the second slider 408 using a belt 411 connected to the cassette holder 409, and a motor 412 driving the belt 411.

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Abstract

The present invention provides an apparatus for isolating nucleic acid or biological material from biological sample using a solid material that carries or absorbs nucleic acid or biological material, the apparatus comprising: a cassette in which plural chambers and input holes into the respective chambers are formed and the plural chambers are connected through a slot which is formed in the bottom of the chambers; a slider which is fitted through the slot to separate the spaces of the chambers, thereby the plural chambers contains a liquid reactant separately, and the slider can be pulled back and forth according to the length of the slot; a cavity which is formed on the chamber side of the slider to settle or collect a solid material in the chamber; a means for pulling the slider to transfer the solid material from one chamber to the next chamber; and a means for agitating the liquid reactants and the solid material in the plural chambers.

Description

APPARATUS FOR ISOLATING NUCLEIC ACID OR BIOLOGICAL MATERIAL
FIELD OF THE INVENTION
The present invention relates to an apparatus for isolating and purifying nucleic acid, cell and various biological materials.
BACKGROUND OF THE INVENTION
Isolation of nucleic acid or biological materials from various biological samples is important starting step in the fields of many areas such as biology, biochemistry, molecular biology, forensic medicine, medical diagnostic and etc. Traditional method for isolating nucleic acid involves harmful organic solvents such as phenol and chloroform. (Sambrook, J., E. F. Fritsoh and T. Maniatis 1989. Molecular cloning: A laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor. N. Y.) Recently, several methods were provided using materials that have the proclivity of binding nucleic acid. Concrete examples of these materials are silica (Boom, R., Sol, C. J. A., Salimans, M. M. M., Jansen, C. L., Wertheim-van Dillen, P. M. E., and van der Noordaa, J. (1990) J. Clin, Microbiol. 28, 495-503), glass fibers (Vogelstein, B., and Gillespie, D. (1979) Proc. Natl. Acad. Sci. USA 76, 615-619, Marko, M. A., Chlpperfield, R., and Birnbolm, H. C. ( 1982) Anal. Biochem. 121 , 382- 387), anion exchange resins (Wang, K., Gan, L., Boysen, C, and Hood, L. (1995) Anal. Biochem. 226, 85-90) and modified magnetic beads (Rudi, K., Kroken, M.s Dahlherg, O. J., Deggerdal, A., Jakobsen, K. S ., and Larsen, F. (1997) BioTechniques 22, 506-51 1 , and Deggerdal, A., and Larsen, F. (1997) BioTechniques 22, 554-557).
The advantages of the method using these materials is that no harmful organic solvent is involved, and physical and biochemical degrading of nucleic acid during the isolation process is minimized. In addition, immobilized nucleic acid is less susceptible to digestion by nucleic acid degrading enzymes.
The above mentioned methods, however, still need intensive manual pipetting steps to transfer the solid material to other vessel and thus the performer is vulnerable to potential viral and bacterial infection from infectious virus and bacteria if infected blood or bacteria is starting material of nucleic acid isolation.
In order to avoid the intensive and tedious manual steps and to eliminate performer 's potential error, several automatic machines such as "MagNa Pure LC" (Roche, Mannheim, Germany AG), "GENESIS" (Techan, Hombrechtikon, Switzerland) and others were developed for high number of sample manipulation. Most of them apply magnetic bead to collect nucleic acid or biological materials from various biological samples to eliminate the use of harmful chemical solvents and centrifugation step. Although these machines are adequate to continuous isolation of nucleic acid or biological material and secure high throughput, they are expensive, rather complicate and inefficient for low and medium number of sample manipulation so that they are not practical for most research and clinical laboratories.
In light of these drawbacks of the traditional apparatus for isolating nucleic acid or biological material, there is a need for an apparatus that is efficient, inexpensive and reproducible as well as applicable to various biological samples.
SUMMARY OF THE INVENTION
A feature of the present invention is to provide an apparatus for isolating a nucleic or biological material from low and medium number of biological samples in which intensive manual pipetting step is eliminated and the risk of viral and bacterial infections from the biological samples is reduced.
Another feature of the present invention is to provide an apparatus for isolating nucleic acid or biological material from the biological samples less expensive and more readily available to researchers, medical personnel and other potential users of the technology.
According to the one aspect of the present invention, there is provided an apparatus for isolating nucleic acid or biological material from biological sample using a solid material which carries or absorbs nucleic acid or biological material, the apparatus comprising:
(a) a cassette in which plural chambers and input holes into the respective chambers are formed and the plural chambers are connected through a slot which is formed in the bottom of the chambers;
(b) a slider which is fitted through the slot to separate the spaces of the chambers, thereby the plural chambers contains a liquid reactant separately, and the slider can be pulled back and forth according to the length of the slot,;
(c) a cavity which is formed on the chamber side of the slider to settle or collect a solid material in the chamber;
(d) a means for pulling the slider to transfer the solid material from one chamber to the next chamber; and
(e) a means for agitating the liquid reactants and the solid material in the plural chambers.
According to the another aspect of the present invention, there is provided an apparatus for isolating nucleic acid or biological material from biological sample using a solid material which carries or absorbs nucleic acid or biological material, the apparatus comprising:
(f) a cassette in which plural chambers and input holes into the respective chambers are formed, the bottoms of the plural chambers are connected through a first slot and the tops of the plural chambers are connected through a second slot;
(g) a first slider which is fitted through the first slot to separate the spaces of the chambers, thereby the plural chambers contains a liquid reactant separately, and the first slider can be -pulled back and forth according to the length of the first slot,; (h) a cavity which is formed on the chamber side of the first slider to settle or collect a solid material in the chamber;
(i) a second slider having plural intruders toward the plural chamber to agitate the liquid reactants and the solid material, which is fitted through the second slot and can be pulled back and forth according to the length of the second slot;
(j) a means for pulling the first slider to transfer the solid material from one chamber to the next chamber; (k) a means for pulling the second slider to agitate the liquid reactants and the solid material; and
(1) a means for agitating the liquid reactants and the solid material in the plural chambers. In the present invention, depending on the kinds of solid material, the various biological samples can be applied over isolation of nucleic acid or biological materials. The various biological samples are blood, bacteria, cultured cells, polymerase chain Reaction (PCR) samples, DNA sequencing samples, DNA-containing agarose product and other biological reaction samples.
In the present invention, the examples of the solid material absorbing or binding nucleic acid or biological materials are silica, glass fiber, anion exchange resins, natural magnetic bead, modified magnetic bead, Sephadex, Sepharose and Sephacryl. Some solid material such as silica, glass fibers, anion exchange resins, Sephadex, Sephacryl and Sepharose have the binding ability to nucleic acid or biological material and can be settled down somewhat rapidly under gravity. The other solid material such as natural or modified magnetic bead have the binding ability to nucleic acid or biological materials depending on modification on surface and affinity to magnet. The feature of the solid material allows the collection of the solid material into the cavity of the slider.
The advantages over the conventional isolation of nucleic acid or biological materials are: i) There is no need for tedious manual pipetting steps and therefore it is possible to eliminate the potential human error such as contamination and mishandling of the samples during the process; ii) The risk of infection by potential infectious agents inside the sample such as virus and bacteria is greatly reduced since the isolation procedure is done in a closed cassette; and iii) The number of samples analyzed at one time can be varied depending on the sets of chamber inside a cassette.
BRIEF DESCRIPTION OF THE DRAWINGS
The above and other obj ect, feature, and advantage of the present invention will be apparent from the following detailed description of the preferred embodiments of the invention in conjunction with the accompanying drawings:
FIG. 1 is a cross sectional view illustrating sequential steps of isolating a nucleic or biological materials using an apparatus according to the present invention;
FIG. 2 is a top view of an apparatus according to the present invention in which a cassette contains eight sets of plural chambers;
FIG. 3 is a cross sectional view of an apparatus according to one example of the present invention; and
FIG. 4 is a cross sectional view of an apparatus according to another example of the present invention.
DESCRIPTION OF THE PREFERRED EMBODIMENT
The present invention provides an apparatus for isolating nucleic acid or biological material from various biological samples. The feature in the present invention is that solid material such as silica, glass fibers, exchange resin, modified magnetic beads and etc. is used for binding or absorbing nucleic acid or biological materials to remove the use of organic solvents and the centrifugation step. To eliminate the pipetting steps, a slider is applied for efficient movement of solid material binding nucleic acid or biological materials.
FIG. 1 is a cross sectional view illustrating sequential steps of isolating a nucleic or biological materials using an apparatus according to a preferred embodiment of the present invention. As shown in FIG. 1, a cassette 100 comprises four chambers 102a,
102b, 102c and 102d with respective input holes 101a, 101b, 101c and lOld. The four chambers 102a, 102b, 102c and 102d are connected with one another with a slot which is fitted with a slider 105. Although the slider 105 can be pulled back and forth according to the length of the slot, the slider 105 fits into the slot very tightly so that the spaces of the chambers 102a, 102b, 102c and 102d are separated and the liquid reactants 103a, 103b, 103c and 103d which is added into the chambers 102a, 102b, 102c and 102d are sustained respectively. The slider 105 has a small cavity 106 that is capable to settle or collect a solid material at one of its end parts. The depth, area and shape of the cavity can be varied depending on the kind of used solid material and the shape of the chambers 102a, 102b, 102c and 102d. Preferably, the cross sectional shape of the cavity 106 is the same with that of the chambers 102a, 102b, 102c and 102d.
The liquid reactants 103a, 103b, 103c and 103d in each chambers 102a, 102b, 102c and 102d can be various buffers such as lysis buffer, ethanol elution buffer, distilled water and etc. which is needed in the process of isolating nucleic acid or biological materials.
In case of isolating nucleic acid from blood using magnetic bead as solid material, the process starts with an addition of blood into the chamber 102a that already contains lysis buffer 103a and a magnetic bead 104. To facilitate the lysis reaction, the whole cassette 100 is shaken, for example, back and forth, or in seesawing or rotating motion in conjunction with motioning system (not shown). After shaking of the cassette 100, a magnet 107 is applied to collect the DNA-carrying magnetic bead 104 into the cavity 106 of the slider 105 as shown in FIG.
1(a). To transfer the magnetic bead 104 to the next chamber 102b, the slider 105 is pulled manually or by automatic pulling system driven by a motor (not shown) and the cavity 106 is located in the bottom of the chamber 102b as shown in FIG. 1(b). After that, for washing the magnetic bead 104, the whole cassette
100 is shaken, for example, back and forth or in seesawing or rotating motion for agitating the liquid reactants 103b and the magnetic bead 104, and then the magnet 107 is applied to collect the solid material 104. The procedure of pulling the slider 105, shaking of the cassette 100 and applying of the magnet 107 is repeated in the step of FIG. 1(c) and (d).
In FIG. 1(d), the liquid reactant 103d is an elution buffer to elute the nucleic acid from magnetic bead 104 and finally the liquid reactant 103d containing nucleic acid in the last chamber 102d is collected by pipetting through an input hole lOld. Although the input hole 101a, 101b, 101c and lOld was shown up in a simplified format in FIG. 1 , other input methods such as a self-sealing piercable diaphragm, and etc. can be used. Also the input holes 101a, 101b, 101c and lOld can be equipped with a sliding lid to avoid flowing of the liquid reactants 103a, 103b, 103c and 103d. In FIG. 1, the step of magnet 107 application to collect the magnetic bead 104 can be omitted if the other type of solid materials such as silica and resins that are easily settled down by gravity is used instead of the magnetic bead 104.
In FIG. 2, a cassette 201 contains eight sets of the chambers 202 for isolating nucleic acid or biological materials from eight different samples. A slider 205 can be pulled according to the direction of the arrow. Although the eight sets of the chambers are shown, the number of the chamber set can be varied from 1 to 20,000. The number of chamber set can be increased by preferably reducing the size of each chamber or enlarging the size of the cassette. The shape of each chamber can be varied within limited area of the chamber.
FIG. 3 is a cross sectional view of an apparatus according to one embodiment of the present invention.
In the present embodiment, the apparatus is explained focused on a means for pulling the slider, a means for generating magnetic force, and a means for agitating the liquid reactants and the solid material.
Agitation of the liquid reactants and the solid material is achieved by a seesawing part 311 that holds and seesaws the cassette.
The means for pulling the slider comprises a belt 314, a hook 313 that fixes the slider to the belt 314, a pivoting point 312 to hold one side of the belt 314, and a motor 315 that drives the belt 314. Accordingly, the slider is pulled by driving the motor 315.
The means for generating magnetic force comprises a magnet 318, a belt 317, a pivoting point 316 to hold one side of the belt 317 and a motor 319 to drives the belt. Thereby, a magnet can be moved according to the movement of magnetic solid material.
Addition to that, a means for controlling the temperature of the plural chambers was added. The means for controlling the temperature is a heater 320 which is operated by electrical power and controlled by a censor 321.
These four parts can be coordinated by programmable software to prosecute the isolation of nucleic acid or biological materials appropriately. FIG. 4 is a cross sectional view of an apparatus according to the other embodiment of the present invention. The main difference of the apparatus in FIG. 4 from the apparatus in FIG. 3 is addition of a second slider 408 on the upper part of a cassette 400 instead of seesawing part, and a system to pull the cassette body 400 and a cassette holder 408 instead of the system to pull a slider 405 and a magnet 407. To simplify the figure, the input holes are omitted. The second slider 408 is a flat plate having multi number of vertical intruder 410. The purpose of the second slider 408 is to agitate the liquid reactants and the solid material in the plural chambers. The second slider 408 has an advantage especially for the case that the mixing reaction is well not achieved by shaking the cassette since the liquid reactants have high surface tension. The liquid reactants and the solid material are agitated by pulling the second slider 408 back and forth repeatedly. A means for pulling the second slider 408 comprises a belt (not shown), a hook 413 that fixes the slider to the belt, and a motor (not shown) that drives the belt.
Movement of the solid material on the cavity is achieved by moving the cassette 400, the cassette holder 409, and the second slider 408 using a belt 411 connected to the cassette holder 409, and a motor 412 driving the belt 411.

Claims

What I claim is;
1 . An apparatus for isolating nucleic acid or biological material from biological sample using a solid material which carries or absorbs nucleic acid or biological material, the apparatus comprising: (a) a cassette in which plural chambers and input holes into the respective chambers are formed and the plural chambers are connected through a slot which is formed in the bottom of the chambers;
(b) a slider which is fitted through the slot to separate the spaces of the chambers, thereby the plural chambers contains a liquid reactant separately, and the slider can be pulled back and forth according to the length of the slot;
(c) a cavity which is formed on the chamber side of the slider to settle or collect a solid material in the chamber; (d) a means for pulling the slider to transfer the solid material from one chamber to the next chamber; and (e) a means for agitating the liquid reactants and the solid material in the plural chambers.
2. The apparatus according to the claim 1 , wherein the slider is in a shape of flat plate in which the cavity is formed at one of its end parts.
3. The apparatus according to the claim 1 , wherein the solid material is selected from the group consisting of silica, glass fiber, anion exchange resins, Sephadex, Sepharose, Sephacryl and magnetic bead.
4. The apparatus according to the claim 1 , further comprising a means for generating magnetic force which is located below the slider to settle or collect a magnetic solid material.
5. The apparatus according to the claim 1 , further comprising a means for controlling a temperature of the plural chambers.
6. The apparatus according to the claim 1 , wherein the operation of the whole apparatus is automated.
7. The apparatus according to the claim 1 , wherein the agitation of the liquid reactants and the solid material is achieved by shaking the cassette in the seesawing motion.
8. An apparatus for isolating nucleic acid or biological material from biological sample using a solid material which carries or absorbs nucleic acid or biological material, the apparatus comprising:
(a) a cassette in which plural chambers and input holes into the respective chambers are formed, the bottoms of the plural chambers are connected through a first slot and the tops of the plural chambers are connected through a second slot;
(b) a first slider which is fitted through the first slot to separate the spaces of the chambers, thereby the plural chambers contains a liquid reactant separately, and the first slider can be pulled back and forth according to the length of the first slot;
(c) a cavity which is formed on the chamber side of the first slider to settle or collect a solid material in the chamber;
(d) a second slider having plural intruders toward the plural chambers to agitate the liquid reactants and the solid material, which is fitted through the second slot and can be pulled back and forth according to the length of the second slot;
(e) a means for pulling the first slider to transfer the solid material from one chamber to the next chamber;
(f) a means for pulling the second slider to agitate the liquid reactants and the solid material; and
(g) a means for agitating the liquid reactants and the solid material in the plural chambers.
9. The apparatus according to the claim 1 , wherein the first slider is in a shape of flat plate in which the cavity is formed at one of its end parts.
10. The apparatus according to the claim 1 , wherein the solid material is selected from the group consisting of silica, glass fiber, anion exchange resins, Sephadex, Sepharose, Sephacryl and magnetic bead.
1 1 . The apparatus according to the claim 1 , further comprising a means for generating magnetic force which is located below the slider to settle or collect a magnetic solid material.
12. The apparatus according to the claim 1 , further comprising a means for controlling a temperature of the plural chambers.
13. The apparatus according to the claim 1 , wherein the operation of the whole apparatus is automated.
14. The apparatus according to the claim 1 , wherein the agitation of the liquid reactants and the solid material is achieved by pulling back and forth according to the length of the second slot.
PCT/KR2001/000153 2001-02-03 2001-02-03 Apparatus for isolating nucleic acid or biological material WO2002063042A1 (en)

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Publication number Priority date Publication date Assignee Title
WO2007097740A3 (en) * 2005-02-11 2007-11-01 Massachusetts Inst Technology Sample preparation methods and devices
US7718072B2 (en) 2002-04-26 2010-05-18 Abbott Laboratories Structure and method for handling magnetic particles in biological assays
WO2016196875A1 (en) * 2015-06-05 2016-12-08 Avanbio Inc. A component of a device, a device, and a method for purifying and testing biomolecules from biological samples
WO2017145080A1 (en) * 2016-02-23 2017-08-31 Bigtec Private Limited A cartridge for purifying a sample and analysis
CN111440706A (en) * 2020-05-25 2020-07-24 烟台澳斯邦生物研发有限公司 A fully enclosed sample nucleic acid extraction and purification device and method

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WO1997008547A1 (en) * 1995-08-24 1997-03-06 Theobald Smith Research Institute, Inc. Method and apparatus for isolating nucleic acid
EP0933132A2 (en) * 1998-02-02 1999-08-04 Toyo Boseki Kabushiki Kaisha Nucleic Acid Extraction Apparatus

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EP0487028A2 (en) * 1990-11-22 1992-05-27 Shimadzu Corporation Apparatus for extracting and purifying nucleic acid
WO1997008547A1 (en) * 1995-08-24 1997-03-06 Theobald Smith Research Institute, Inc. Method and apparatus for isolating nucleic acid
EP0933132A2 (en) * 1998-02-02 1999-08-04 Toyo Boseki Kabushiki Kaisha Nucleic Acid Extraction Apparatus

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7718072B2 (en) 2002-04-26 2010-05-18 Abbott Laboratories Structure and method for handling magnetic particles in biological assays
US8211301B2 (en) 2002-04-26 2012-07-03 Abbott Laboratories Structure and method for handling magnetic particles in biological assays
US8728311B2 (en) 2002-04-26 2014-05-20 Abbott Laboratory Structure and method for handling magnetic particles in biological assays
WO2007097740A3 (en) * 2005-02-11 2007-11-01 Massachusetts Inst Technology Sample preparation methods and devices
WO2016196875A1 (en) * 2015-06-05 2016-12-08 Avanbio Inc. A component of a device, a device, and a method for purifying and testing biomolecules from biological samples
US20220176370A1 (en) * 2015-06-05 2022-06-09 The Emerther Company Method for purifying and testing biomolecules from biological samples
WO2017145080A1 (en) * 2016-02-23 2017-08-31 Bigtec Private Limited A cartridge for purifying a sample and analysis
CN111440706A (en) * 2020-05-25 2020-07-24 烟台澳斯邦生物研发有限公司 A fully enclosed sample nucleic acid extraction and purification device and method

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