US20030199078A1 - Method, kit and apparatus for the isolation of nucleic acids - Google Patents
Method, kit and apparatus for the isolation of nucleic acids Download PDFInfo
- Publication number
- US20030199078A1 US20030199078A1 US10/426,641 US42664103A US2003199078A1 US 20030199078 A1 US20030199078 A1 US 20030199078A1 US 42664103 A US42664103 A US 42664103A US 2003199078 A1 US2003199078 A1 US 2003199078A1
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- US
- United States
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- sample
- analyte
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- Abandoned
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- 238000000034 method Methods 0.000 title claims abstract description 79
- 108020004707 nucleic acids Proteins 0.000 title claims abstract description 21
- 102000039446 nucleic acids Human genes 0.000 title claims abstract description 21
- 150000007523 nucleic acids Chemical class 0.000 title claims abstract description 21
- 238000002955 isolation Methods 0.000 title claims abstract description 17
- 239000000523 sample Substances 0.000 claims abstract description 102
- 238000002360 preparation method Methods 0.000 claims abstract description 46
- 239000012491 analyte Substances 0.000 claims abstract description 45
- 239000011521 glass Substances 0.000 claims abstract description 40
- 239000002245 particle Substances 0.000 claims abstract description 31
- 230000005291 magnetic effect Effects 0.000 claims abstract description 20
- 239000012472 biological sample Substances 0.000 claims abstract description 14
- 239000000725 suspension Substances 0.000 claims abstract description 10
- 238000006243 chemical reaction Methods 0.000 claims description 58
- 238000001179 sorption measurement Methods 0.000 claims description 53
- 239000011159 matrix material Substances 0.000 claims description 46
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 31
- 238000002156 mixing Methods 0.000 claims description 24
- 238000010828 elution Methods 0.000 claims description 19
- 229910011255 B2O3 Inorganic materials 0.000 claims description 15
- 239000003153 chemical reaction reagent Substances 0.000 claims description 15
- 229910052681 coesite Inorganic materials 0.000 claims description 15
- 229910052906 cristobalite Inorganic materials 0.000 claims description 15
- JKWMSGQKBLHBQQ-UHFFFAOYSA-N diboron trioxide Chemical compound O=BOB=O JKWMSGQKBLHBQQ-UHFFFAOYSA-N 0.000 claims description 15
- 230000002934 lysing effect Effects 0.000 claims description 15
- 239000000377 silicon dioxide Substances 0.000 claims description 15
- 229910052682 stishovite Inorganic materials 0.000 claims description 15
- 229910052905 tridymite Inorganic materials 0.000 claims description 15
- 238000010438 heat treatment Methods 0.000 claims description 13
- 239000007787 solid Substances 0.000 claims description 12
- 239000011534 wash buffer Substances 0.000 claims description 11
- PNEYBMLMFCGWSK-UHFFFAOYSA-N aluminium oxide Inorganic materials [O-2].[O-2].[O-2].[Al+3].[Al+3] PNEYBMLMFCGWSK-UHFFFAOYSA-N 0.000 claims description 9
- ODINCKMPIJJUCX-UHFFFAOYSA-N calcium oxide Inorganic materials [Ca]=O ODINCKMPIJJUCX-UHFFFAOYSA-N 0.000 claims description 9
- 229910052593 corundum Inorganic materials 0.000 claims description 9
- 229910001845 yogo sapphire Inorganic materials 0.000 claims description 9
- 108010067770 Endopeptidase K Proteins 0.000 claims description 8
- 230000003321 amplification Effects 0.000 claims description 8
- 238000003199 nucleic acid amplification method Methods 0.000 claims description 8
- 238000005406 washing Methods 0.000 claims description 8
- KKCBUQHMOMHUOY-UHFFFAOYSA-N Na2O Inorganic materials [O-2].[Na+].[Na+] KKCBUQHMOMHUOY-UHFFFAOYSA-N 0.000 claims description 7
- 108091005804 Peptidases Proteins 0.000 claims description 7
- 239000000872 buffer Substances 0.000 claims description 7
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- 239000004365 Protease Substances 0.000 claims description 6
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims description 6
- 238000001816 cooling Methods 0.000 claims description 6
- 235000019419 proteases Nutrition 0.000 claims description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- 239000000203 mixture Substances 0.000 claims description 5
- 102000004190 Enzymes Human genes 0.000 claims description 4
- 108090000790 Enzymes Proteins 0.000 claims description 4
- 239000013578 denaturing buffer Substances 0.000 claims description 4
- 238000007885 magnetic separation Methods 0.000 claims description 4
- NOTVAPJNGZMVSD-UHFFFAOYSA-N potassium monoxide Inorganic materials [K]O[K] NOTVAPJNGZMVSD-UHFFFAOYSA-N 0.000 claims description 4
- ZRALSGWEFCBTJO-UHFFFAOYSA-N Guanidine Chemical class NC(N)=N ZRALSGWEFCBTJO-UHFFFAOYSA-N 0.000 claims description 3
- 229910000272 alkali metal oxide Inorganic materials 0.000 claims description 3
- 229910000287 alkaline earth metal oxide Inorganic materials 0.000 claims description 3
- PJJJBBJSCAKJQF-UHFFFAOYSA-N guanidinium chloride Chemical compound [Cl-].NC(N)=[NH2+] PJJJBBJSCAKJQF-UHFFFAOYSA-N 0.000 claims description 3
- 150000003839 salts Chemical class 0.000 claims description 3
- ZJYYHGLJYGJLLN-UHFFFAOYSA-N guanidinium thiocyanate Chemical compound SC#N.NC(N)=N ZJYYHGLJYGJLLN-UHFFFAOYSA-N 0.000 claims description 2
- 239000003960 organic solvent Substances 0.000 claims description 2
- 238000001514 detection method Methods 0.000 abstract description 10
- 238000004519 manufacturing process Methods 0.000 abstract description 2
- 108020004414 DNA Proteins 0.000 description 10
- 239000012071 phase Substances 0.000 description 10
- 239000006249 magnetic particle Substances 0.000 description 8
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 7
- 241000606161 Chlamydia Species 0.000 description 7
- 230000009089 cytolysis Effects 0.000 description 7
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- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 6
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- 239000007788 liquid Substances 0.000 description 5
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- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 4
- 239000006148 magnetic separator Substances 0.000 description 4
- 239000011148 porous material Substances 0.000 description 4
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- 239000008280 blood Substances 0.000 description 3
- 230000003196 chaotropic effect Effects 0.000 description 3
- 229940088598 enzyme Drugs 0.000 description 3
- 210000002381 plasma Anatomy 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 238000003860 storage Methods 0.000 description 3
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 2
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 2
- 101100203596 Caenorhabditis elegans sol-1 gene Proteins 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 2
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- 102000004169 proteins and genes Human genes 0.000 description 2
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- 102000003960 Ligases Human genes 0.000 description 1
- 108090000364 Ligases Proteins 0.000 description 1
- 102000016943 Muramidase Human genes 0.000 description 1
- 108010014251 Muramidase Proteins 0.000 description 1
- 108010062010 N-Acetylmuramoyl-L-alanine Amidase Proteins 0.000 description 1
- 108010067372 Pancreatic elastase Proteins 0.000 description 1
- 102000016387 Pancreatic elastase Human genes 0.000 description 1
- 102000035195 Peptidases Human genes 0.000 description 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- 108010059712 Pronase Proteins 0.000 description 1
- -1 RNases Chemical class 0.000 description 1
- 102000006382 Ribonucleases Human genes 0.000 description 1
- 108010083644 Ribonucleases Proteins 0.000 description 1
- 101150058348 SWE1 gene Proteins 0.000 description 1
- 238000002105 Southern blotting Methods 0.000 description 1
- 239000007983 Tris buffer Substances 0.000 description 1
- 229920004890 Triton X-100 Polymers 0.000 description 1
- 108020005202 Viral DNA Proteins 0.000 description 1
- 108020000999 Viral RNA Proteins 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 230000001476 alcoholic effect Effects 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
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- 239000006166 lysate Substances 0.000 description 1
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- 108020004999 messenger RNA Proteins 0.000 description 1
- 239000002923 metal particle Substances 0.000 description 1
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- 229910052618 mica group Inorganic materials 0.000 description 1
- 239000012299 nitrogen atmosphere Substances 0.000 description 1
- 239000002773 nucleotide Substances 0.000 description 1
- 125000003729 nucleotide group Chemical group 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
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- 229910052760 oxygen Inorganic materials 0.000 description 1
- VLTRZXGMWDSKGL-UHFFFAOYSA-M perchlorate Inorganic materials [O-]Cl(=O)(=O)=O VLTRZXGMWDSKGL-UHFFFAOYSA-M 0.000 description 1
- VLTRZXGMWDSKGL-UHFFFAOYSA-N perchloric acid Chemical compound OCl(=O)(=O)=O VLTRZXGMWDSKGL-UHFFFAOYSA-N 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
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- BDERNNFJNOPAEC-UHFFFAOYSA-N propan-1-ol Chemical compound CCCO BDERNNFJNOPAEC-UHFFFAOYSA-N 0.000 description 1
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- 238000003757 reverse transcription PCR Methods 0.000 description 1
- 230000007017 scission Effects 0.000 description 1
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- 229910052814 silicon oxide Inorganic materials 0.000 description 1
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- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
- 150000003672 ureas Chemical class 0.000 description 1
Images
Classifications
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B03—SEPARATION OF SOLID MATERIALS USING LIQUIDS OR USING PNEUMATIC TABLES OR JIGS; MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
- B03C—MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
- B03C1/00—Magnetic separation
- B03C1/005—Pretreatment specially adapted for magnetic separation
- B03C1/01—Pretreatment specially adapted for magnetic separation by addition of magnetic adjuvants
-
- C—CHEMISTRY; METALLURGY
- C03—GLASS; MINERAL OR SLAG WOOL
- C03C—CHEMICAL COMPOSITION OF GLASSES, GLAZES OR VITREOUS ENAMELS; SURFACE TREATMENT OF GLASS; SURFACE TREATMENT OF FIBRES OR FILAMENTS MADE FROM GLASS, MINERALS OR SLAGS; JOINING GLASS TO GLASS OR OTHER MATERIALS
- C03C14/00—Glass compositions containing a non-glass component, e.g. compositions containing fibres, filaments, whiskers, platelets, or the like, dispersed in a glass matrix
- C03C14/004—Glass compositions containing a non-glass component, e.g. compositions containing fibres, filaments, whiskers, platelets, or the like, dispersed in a glass matrix the non-glass component being in the form of particles or flakes
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
- C07H1/06—Separation; Purification
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
- C07H1/06—Separation; Purification
- C07H1/08—Separation; Purification from natural products
-
- 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
- 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
- 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
- G01N35/0098—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor involving analyte bound to insoluble magnetic carrier, e.g. using magnetic separation
-
- 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
- G01N35/0099—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor comprising robots or similar manipulators
-
- H—ELECTRICITY
- H01—ELECTRIC ELEMENTS
- H01F—MAGNETS; INDUCTANCES; TRANSFORMERS; SELECTION OF MATERIALS FOR THEIR MAGNETIC PROPERTIES
- H01F1/00—Magnets or magnetic bodies characterised by the magnetic materials therefor; Selection of materials for their magnetic properties
- H01F1/01—Magnets or magnetic bodies characterised by the magnetic materials therefor; Selection of materials for their magnetic properties of inorganic materials
- H01F1/03—Magnets or magnetic bodies characterised by the magnetic materials therefor; Selection of materials for their magnetic properties of inorganic materials characterised by their coercivity
- H01F1/032—Magnets or magnetic bodies characterised by the magnetic materials therefor; Selection of materials for their magnetic properties of inorganic materials characterised by their coercivity of hard-magnetic materials
- H01F1/10—Magnets or magnetic bodies characterised by the magnetic materials therefor; Selection of materials for their magnetic properties of inorganic materials characterised by their coercivity of hard-magnetic materials non-metallic substances, e.g. ferrites, e.g. [(Ba,Sr)O(Fe2O3)6] ferrites with hexagonal structure
- H01F1/11—Magnets or magnetic bodies characterised by the magnetic materials therefor; Selection of materials for their magnetic properties of inorganic materials characterised by their coercivity of hard-magnetic materials non-metallic substances, e.g. ferrites, e.g. [(Ba,Sr)O(Fe2O3)6] ferrites with hexagonal structure in the form of particles
- H01F1/112—Magnets or magnetic bodies characterised by the magnetic materials therefor; Selection of materials for their magnetic properties of inorganic materials characterised by their coercivity of hard-magnetic materials non-metallic substances, e.g. ferrites, e.g. [(Ba,Sr)O(Fe2O3)6] ferrites with hexagonal structure in the form of particles with a skin
Definitions
- the invention concerns a process for preparing biological samples for the subsequent detection of an analyte, in particular a nucleic acid, in this sample.
- analyte in particular a nucleic acid
- reagent kits and new devices for sample preparation and new magnetic pigments are provided.
- the sample preparation often has to meet special requirements in a method for the detection of an analyte in a biological sample.
- the analyte is often present at a very low concentration and, on the other hand, there are often many other substances in the sample which can interfere with the isolation or determination of the analyte.
- WO 96/41811 discloses a process for the isolation of an analyte, especially a nucleic acid, from a biological sample wherein the sample which contains the analyte in a liquid is contacted with magnetic particles that have an outer glass surface which is essentially free of pores or has pores with a diameter of ⁇ 10 nm, under conditions such that the analyte binds to the particle surface and the bound analyte is separated from the sample liquid.
- the process described in WO 96/46811 is very well suited to the purification of an analyte from a biological sample. However, it cannot be easily applied to an automated sample preparation. Boom et al. (J. Clin. Microbiol. 28 (1990), 495-503) also describe a protocol for the purification of nucleic acids from a biological sample using silicon oxide particles fractionated according to size. However, this process is complicated and not suitable for automation and moreover there is a risk of carry-over.
- the object of the present invention was to provide a new sample preparation process in which the disadvantages of the state of the art are at least partially eliminated.
- a further aspect of the present invention is a process for the isolation of an analyte from a biological sample comprising the steps:
- the process according to the invention is based on the selective binding of analytes to a solid adsorption matrix in the presence of a sample lysing buffer in which the analyte that is preferably a nucleic acid such as DNA e.g. chromosomal DNA, fragmented chromosomal DNA, plasmid DNA, viral DNA etc. or RNA e.g. mRNA, tRNA, rRNA or viral RNA etc., is separated from impurities of the sample such as proteins or cell debris.
- the sample can be any biological sample e.g. a body fluid such as blood, plasma, urine etc., a tissue sample, a sample of cultured cells or such like.
- the adsorption matrix used in the process according to the invention is able to ensure the substantially selective binding of the analyte under the reaction conditions.
- a particulate adsorption matrix is preferably used which preferably contains a glass surface.
- Magnetic glass particles are particularly preferred, especially the magnetic particles described in WO 96/41811 with an external glass surface which is essentially free of pores or has pores with a diameter of less than 10 nm.
- Ferromagnetic particles are particularly preferred which have a particle size between 10 and 60 ⁇ m.
- Such particles can for example contain a core made of mica and magnetic particles immobilized thereon which is enclosed by a layer of glass. Whereas in WO 96/41811 the magnetic particles are placed in the individual reaction vessels in a solid form e.g.
- the magnetic particles are preferably used according to the invention in the form of a suspension.
- Alcoholic suspensions having a concentration of about 5 to 20 mg/ml have proven to be particularly suitable. It was surprisingly found that, despite the high specific density of the magnetic glass particles, the suspension can be very reproducibly drawn out of a storage container which enables the process to be automated.
- glass particles described in WO 96/41811 give good results in the process according to the invention, particularly good results are obtained with glass particles whose glass phase contains the following metal oxides: SiO 2 , B 2 O 3 , alkali metal oxide e.g. K 2 O or/and Na 2 O and optionally Al 2 O 3 and an alkaline earth metal oxide e.g. CaO.
- metal oxides SiO 2 , B 2 O 3 , alkali metal oxide e.g. K 2 O or/and Na 2 O and optionally Al 2 O 3 and an alkaline earth metal oxide e.g. CaO.
- the contents of these metal oxides are preferably as follows: 50 to 95 mol-% SiO 2 , 0.2 to 30 mol-% B 2 O 3 , 0 to 10 mol-% Al 2 O 3 , 0 to 20 mol-% alkaline earth metal oxide and 0.2 to 20 mol-% alkali metal oxide where the percentages are each based on the total weight of the glass phase.
- a glass phase which contains SiO 2 , B 2 O 3 , K 2 O, Al 2 O 3 and CaO has proven to be particularly suitable for the isolation of RNA.
- a glass phase which contains SiO 2 , B 2 O 3 and Na 2 O has proven to be particularly suitable for the isolation of DNA.
- the adsorption matrix is preferably added in an amount which corresponds to the minimum amount required to quantitatively bind the analyte present in the sample, in particular a nucleic acid, or the amount is somewhat larger, preferably at most 50% and particularly preferably at most 20% above this amount.
- the expected amount of nucleic acid in various type of samples can—if it is not already known—be determined in advance by common techniques e.g. phenol/chloroform extraction and subsequent measurement of the optical density.
- Step (a) of the process according to the invention comprises lysing the sample in a reaction vessel.
- This lysis is usually carried out by lysing the cells present in the sample under denaturing conditions e.g. by adding a protease and a denaturing buffer.
- Proteinase K, pronase, elastase or/and lysozyme are preferably used as the proteinase. The use of proteinase K is particularly preferred.
- the protease digestion is carried out in a denaturing buffer which contains a chaotropic compound e.g. urea or urea derivatives, preferably a chaotropic salt,—particularly preferably a guanidinium salt such as guanidinium hydrochloride (especially for the isolation of DNA) or guanidinium thiocyanate (especially for the isolation of RNA) or a perchlorate or iodide. Concentrations in the range of 1 to 3 mol/l are preferred for guanidinium salts.
- a chaotropic compound e.g. urea or urea derivatives, preferably a chaotropic salt,—particularly preferably a guanidinium salt such as guanidinium hydrochloride (especially for the isolation of DNA) or guanidinium thiocyanate (especially for the isolation of RNA) or a perchlorate or iodide.
- the solid adsorption matrix is only added after lysing the sample. This procedure results in a significantly lower unspecific binding of undesired sample components, e.g. proteins, to the adsorption matrix.
- step (c) the analyte is selectively bound to the adsorption matrix by incubation in the lysing buffer preferably under chaotropic conditions.
- Step (d) of the process according to the invention comprises the separation of non-bound sample components from the adsorption matrix.
- the non-bound sample components are preferably removed from the reaction vessel. This can be achieved by adding and removing a wash buffer, optionally several times, which preferably contains a quantity of at least 50% (v/v) and particularly preferably of at least 60% (v/v) of a solvent that is miscible with water such as ethanol, propanol and acetone.
- Steps (c), (d) or/and (e) of the process according to the invention are preferably carried out while mixing continuously or at intervals (i.e. mixing phases alternate with phases in which the reaction vessel is at rest) without adding external means.
- This mixing is preferably carried out by rotating the reaction vessel around its longitudinal axis while reversing the direction of rotation several times.
- the mixing vessel is particularly preferably rotated exactly around its longitudinal axis and the change in the direction of rotation is carried out such that the meniscus deflection of the liquid remains below ⁇ a predetermined cut-off value.
- Such mixing processes are described in WO 91/15768 and EP-A-0 435 481.
- the duration of steps (c) or/and (e) is preferably 20 min at most and comprises a continuous mixing or an interval mixing in short cycles, preferably in short cycles of preferably two minutes maximum. Particularly good results were obtained by interval mixing in a one minute cycle comprising 20 sec mixing and 40 sec resting.
- Step (e) of the process according to the invention comprises the elution of the analyte from the adsorption matrix.
- a low salt buffer that is essentially free of organic solvents can be used for this as is known from the prior art.
- the elution buffer can contain additional reagents such as enzymes e.g. enzymes used to manipulate nucleic acids such as RNases, DNases, restriction endonucleases, ligases, terminal transferases or/and polymerases. If the analyte is for example a DNA it is possible to add a DNase-free RNase during the elution in order to reduce the content of undesired RNA.
- analyte is RNA
- RNase-free DNase it is possible to add an RNase-free DNase during the elution.
- Other enzymes such as restriction endonucleases etc. can be added in an analogous manner.
- a nucleic acid amplification master mix which contains the amplification buffer, nucleotides, primers, polymerase and buffer salts can also be added during the elution.
- Step (f) of the process according to the invention comprises separating the eluate from the adsorption matrix. This separation can be carried out in the usual manner e.g. by sedimentation but preferably by magnetic separation.
- the analytes isolated by the process according to the invention can be subsequently processed further in a known manner e.g. in the case of nucleic acids by amplification and subsequent detection, or detection without previous amplification or sequencing.
- various analytes can be determined in aliquots of the eluate e.g. various viruses such as HIV, HCV and HBV.
- An important feature of the process according to the invention is that many or optionally even all steps can be carried out at essentially the same temperature i.e. within a temperature range of ⁇ 2.5° C.
- This temperature is preferably in the range of room temperature to 70° C., particular preferably from room temperature to 40° C., most preferably at room temperature i.e. ca. 18 to 32° C.
- at least the steps (c) of adsorption and (d) of washing are carried out at this temperature.
- Other steps, in particular the steps (a) of lysing or/and (e) of elution are particularly preferably also carried out at this temperature.
- the entire sample preparation can for example be carried out at a uniform temperature for the determination of HIV in blood samples.
- an additional after-treatment step at an elevated temperature can take place after step (f) of the process according to the invention which improves the amplification yields for certain analytes. It may be necessary for other analytes to carry out the pre-treatment or/and the elution at an elevated temperature.
- the elevated temperature is preferably in the range of more than 40° C. to 95° C. e.g. ca. 70° C.
- the process according to the invention is preferably carried out in an automated device. Examples of such devices are described in the following. It is also preferable that in the process according to the invention for sample preparation at least steps (a) to (e) are carried out in a single reaction vessel i.e. that there is no transfer into another reaction vessel. This considerably simplifies the process and also leads to a reduction of the risk of contamination.
- reagent kit which is especially suitable for carrying out the process described above comprising
- reagent kit for isolating DNA comprising magnetic glass particles whose glass phase contains SiO 2 , B 2 O 3 and Na 2 O
- reagent kit for isolating RNA comprising magnetic glass particles whose glass phase contains SiO 2 , B 2 O 3 , Al 2 O 3 , CaO and Ka 2 O.
- Another subject matter of the present invention is a device for isolating an analyte from a biological sample comprising:
- a first holding device for reaction vessels for sample preparation ( 3 ) which is equipped for an operating temperature of ⁇ 70° C., in particular ⁇ 40° C.,
- a second holding device for reaction vessels ( 4 a , 4 b , 4 c ), which optionally contains a cooling or/and heating means,
- the device according to the invention is preferably designed such that a single reaction vessel is used to carry out the 4 main steps of sample preparation i.e. lysis of a sample, adsorption of the released analyte e.g. a nucleic acid to a solid adsorption matrix e.g. magnetic glass particles, washing the adsorption matrix and eluting the analyte from the adsorption matrix.
- sample preparation i.e. lysis of a sample
- adsorption of the released analyte e.g. a nucleic acid
- a solid adsorption matrix e.g. magnetic glass particles
- the device is designed such that the first holding device for holding the reaction vessels for sample preparation is used at least for the adsorption of the analyte to the solid adsorption matrix and for washing the adsorption matrix.
- the first holding device is also used for the sample lysis or/and for the elution of the analyte from the adsorption matrix.
- the reaction vessels for the sample preparation have a volume of preferably at least 1 ml e.g. 1-5 ml.
- the second holding device is designed for reaction vessels to store or/and further process the analyte e.g. PCR vessels which usually have a different shape than the reaction vessels used for the sample preparation.
- the reaction vessels for storing or/and additional processing have a volume of preferably up to 500 ⁇ l, e.g. 50-200 ⁇ l.
- the second holding device can contain vessels for reagents which are required to process the sample containing the analyte e.g. a PCR master mix.
- the device according to the invention can be designed such that one or several steps of the sample preparation or/and an after-treatment step can be carried out at an elevated temperature in the second holding device.
- the second holding device can be designed to hold reaction vessels for at least one treatment step which is selected from lysing the sample, eluting the sample from the adsorption matrix and an after-treatment step after elution.
- the first holding device preferably contains means for the magnetic separation.
- the first holding device contains means for mixing the reaction vessels in particular by rotating them around their longitudinal axis. Such means can optionally be provided for the second holding device.
- the robotic tool generally comprises automatic pipetting devices and optionally means for transporting reaction vessels e.g. between the first and second holding device.
- a cap opening and closing unit may be integrated.
- the sample preparation device ( 1 ) contains a holding device for reagents ( 2 ), a holding device for reaction vessels for sample preparation ( 3 ) with the functions mixing and magnetic separation which provides a temperature of preferably ⁇ 40° C. and particularly preferably room temperature.
- the device additionally contains a holding station for further reaction vessels ( 4 a ) e.g. for PCR vessels which has a temperature of 4° C. to room temperature.
- the device additionally contains automated devices for pipetting and handling reaction vessels ( 5 ) which enables movements in an X, Y and Z direction.
- the four main steps of sample preparation i.e.
- lysis, adsorption, washing and elution take place in a single reaction vessel in the first holding device. Eluates are stored and further reagents e.g. PCR master mix are added in the second holding device. For further processing e.g. for a subsequent PCR, the vessels are transferred to an appropriate device e.g. a thermocycler (not shown).
- an appropriate device e.g. a thermocycler (not shown).
- the device contains a second holding device ( 4 b ) which is designed to hold reaction vessels for further processing e.g. PCR vessels and is equipped to set a temperature of 4° C. (cooling the PCR master mix) to 95° C. to heat the eluate after elution from the adsorption matrix.
- a cap counter-heating is preferred to prevent condensation on the cap of the PCR vessels.
- the embodiment of the device according to the invention shown in FIG. 3 is provided with a second holding device for reaction vessels ( 4 c ) which is designed to hold PCR vessels and sample preparation vessels.
- a second holding device for reaction vessels ( 4 c ) which is designed to hold PCR vessels and sample preparation vessels.
- this second holding device cooling e.g. to 40° C. and heating e.g. to 95° C. is possible to heat the lysate or/and the eluate.
- a cap counter-heating is provided to prevent condensation on the cap of reaction vessels.
- the first holding device is designed to set a temperature in the range of ⁇ 70° C.
- the second holding device is—as shown in FIG. 3—suitable for cooling and heating sample processing and sample preparation vessels.
- the devices according to the invention can be used especially in a process as described above.
- FIG. 1 shows a schematic representation of a first embodiment of the device according to the invention
- FIG. 2 shows a schematic representation of a second embodiment of the device according to the invention
- FIG. 3 shows a schematic representation of a third embodiment of the device according to the invention
- FIG. 4 shows the result of a chlamydia detection by PCR using manual and semiautomatic sample preparation
- FIG. 5 shows the result of a chlamydia detection by PCR using semiautomatic sample preparation and various temperature profiles during the sample preparation
- FIG. 6 shows the result of a HIV detection by PCR using manual sample preparation (standard protocol) and semiautomated sample preparation at room temperature.
- sols were prepared as follows:
- sols were subsequently subjected to a spray drying process.
- the powder obtained by the spray drying was subjected to a separation of fines by sedimentation, a temperature treatment under a nitrogen atmosphere (60 l/h volume flow rate) at a heating rate of 1 K/min and kept for one hour at a compaction temperature in the range of 600 to 700° C. Subsequently the oven was cooled to 300° C. and flushed with oxygen for 1 h at this temperature. After cooling to room temperature the magnetic glass particles were removed and sieved through a 50 ⁇ m sieve to separate the coarse material.
- the magnetic glass particles obtained from sol 1 are particularly suitable for the isolation of DNA.
- the glass particles obtained from sol 2 are particularly suitable for the isolation of RNA.
- nucleic acids obtained in this manner can be used directly after the elution for an amplification by PCR, a restriction cleavage or a Southern blot.
- the reaction kit contains:
- binding buffer (4.7 mol/l guanidinium hydrochloride, 10 mmol/l urea, 10 mmol/l Tris HCl, 20% Triton®X-100, pH 5.7
- wash buffer (56% (v/v) ethanol, 20 mmol/l NaCl, 10 mmol/l Tris HCl, pH 7.5)
- kit components are stable and can be stored at room temperature. After dissolving proteinase K in water, the solution should be aliquoted and stored at ⁇ 20° C. The frozen solution is stable for 12 months.
- reaction vessel is placed in a magnetic particle separator (Boehringer Mannheim, Cat. No. 1 641 794) and separated for about 1 min.
- a magnetic particle separator Boehringer Mannheim, Cat. No. 1 641 794
- nucleic acids e.g. DNA obtained in this manner are stable and can be subsequently directly processed further or stored at 4° C.
- microtitre plates e.g. deep well microtitre plates (e.g. Ritter, J. J. Bioanalytic).
- the MGPs are concentrated by transferring the sample to a magnetic separator. After one minute the supernatant is completely removed by pipette.
- 0.5 ml wash buffer is added to the MGPs by pipette.
- the sample is subjected to vortex mixing and then transferred to the magnetic separator.
- the supernatant is removed by pipette after 1 min. The washing procedure is repeated for a further two times.
- FIG. 4 shows a comparison of the determination of chlamydia (sample: 100 elementary antibodies per 100 ml urine; six-fold determination) between the manual standard protocol (vortex) and the semiautomated process (MTM). It can be seen that the sensitivity is not impaired by the automation.
- sample preparation is carried out as described in section 3.2. However, the lysis and elution are carried out at room temperature.
- sample preparation is carried out as described in section 3.3. After elution an incubation is carried out for 10 min at 70° C.
- FIG. 5 shows a comparison of the chlamydia determination (samples: SWE1, O chlamydia elementary antibodies (EAB) per ml urine, SWE2: 10 EAB, SWE3: 100 EAB and SWE 4:1000 EAB each per ml urine) between the sample preparation protocols described in sections 3.2, 3.3 and 3.4.
- EAB O chlamydia elementary antibodies
- SWE2 10 EAB
- SWE3 100 EAB and SWE 4:1000 EAB each per ml urine
- Frozen plasma is thawed for 5 min at 37° C. and cooled on ice for further processing.
- 750 ⁇ l wash buffer is added to the MGPs.
- the MGPs are resuspended and separated as described previously.
- the wash procedure is repeated four times and the washing buffer is carefully removed at the end.
- sample preparation is carried out as described in section 4.1 except that the mixing and heating was carried out on a mixing and heating module.
- sample preparation is essentially carried out as described in section 4.2 except that all steps are carried out at room temperature.
- the incubation period for lysis, adsorption and elution is in each case 15 min.
Abstract
The invention concerns a process for preparing biological samples for the subsequent detection of an analyte. In particular, the invention relates to a process for the isolation of a nucleic acid in a sample using a suspension of magnetic glass particles. In addition, kits and apparatuses containing magnetic glass particles for sample preparation are provided.
Description
- The invention concerns a process for preparing biological samples for the subsequent detection of an analyte, in particular a nucleic acid, in this sample. In addition reagent kits and new devices for sample preparation and new magnetic pigments are provided.
- The sample preparation often has to meet special requirements in a method for the detection of an analyte in a biological sample. On the one hand, the analyte is often present at a very low concentration and, on the other hand, there are often many other substances in the sample which can interfere with the isolation or determination of the analyte.
- WO 96/41811 discloses a process for the isolation of an analyte, especially a nucleic acid, from a biological sample wherein the sample which contains the analyte in a liquid is contacted with magnetic particles that have an outer glass surface which is essentially free of pores or has pores with a diameter of <10 nm, under conditions such that the analyte binds to the particle surface and the bound analyte is separated from the sample liquid. The process described in WO 96/46811 is very well suited to the purification of an analyte from a biological sample. However, it cannot be easily applied to an automated sample preparation. Boom et al. (J. Clin. Microbiol. 28 (1990), 495-503) also describe a protocol for the purification of nucleic acids from a biological sample using silicon oxide particles fractionated according to size. However, this process is complicated and not suitable for automation and moreover there is a risk of carry-over.
- In a method described in EP-A-0 757 106 for the extraction of nucleic acids, a sample is lysed, the nucleic acids present in the sample are bound to superparamagnetic metal particles, these are removed from the sample vessel with a pipette and thus separated from the other sample components. A disadvantage of this method is that losses may occur due to the necessity of having to remove the analyte from the sample with a pipette. Furthermore there is a risk of carry-over and contamination due to the use of several reaction vessels.
- Hence the object of the present invention was to provide a new sample preparation process in which the disadvantages of the state of the art are at least partially eliminated. In particular it should be possible to automate the new process and have a temperature profile that is as simple as possible.
- This object is achieved by a process for the isolation of an analyte from a biological sample comprising the steps:
- (a) lysing the sample in a reaction vessel,
- (b) adding a solid adsorption matrix
- (c) incubating under such conditions that the analyte binds to the adsorption matrix,
- (d) removing non-bound sample components from the reaction vessel,
- (e) incubating under such conditions that the analyte is eluted from the adsorption matrix and
- (f) separating the eluate from the adsorption matrix.
- A further aspect of the present invention is a process for the isolation of an analyte from a biological sample comprising the steps:
- (a) lysing the sample in a reaction vessel,
- (b) adding a solid adsorption matrix
- (c) incubating under such conditions that the analyte binds to the adsorption matrix,
- (d) separating non-bound sample components from the adsorption matrix,
- (e) incubating under such conditions that the analyte is eluted from the adsorption matrix and
- (f) separating the eluate from the adsorption matrix
- wherein at least steps (c) and (d) are carried out at essentially the same temperature.
- The process according to the invention is based on the selective binding of analytes to a solid adsorption matrix in the presence of a sample lysing buffer in which the analyte that is preferably a nucleic acid such as DNA e.g. chromosomal DNA, fragmented chromosomal DNA, plasmid DNA, viral DNA etc. or RNA e.g. mRNA, tRNA, rRNA or viral RNA etc., is separated from impurities of the sample such as proteins or cell debris. The sample can be any biological sample e.g. a body fluid such as blood, plasma, urine etc., a tissue sample, a sample of cultured cells or such like.
- The adsorption matrix used in the process according to the invention is able to ensure the substantially selective binding of the analyte under the reaction conditions. A particulate adsorption matrix is preferably used which preferably contains a glass surface. Magnetic glass particles are particularly preferred, especially the magnetic particles described in WO 96/41811 with an external glass surface which is essentially free of pores or has pores with a diameter of less than 10 nm. Ferromagnetic particles are particularly preferred which have a particle size between 10 and 60 μm. Such particles can for example contain a core made of mica and magnetic particles immobilized thereon which is enclosed by a layer of glass. Whereas in WO 96/41811 the magnetic particles are placed in the individual reaction vessels in a solid form e.g. as tablets or a powder, the magnetic particles are preferably used according to the invention in the form of a suspension. Alcoholic suspensions having a concentration of about 5 to 20 mg/ml have proven to be particularly suitable. It was surprisingly found that, despite the high specific density of the magnetic glass particles, the suspension can be very reproducibly drawn out of a storage container which enables the process to be automated.
- Although the glass particles described in WO 96/41811 give good results in the process according to the invention, particularly good results are obtained with glass particles whose glass phase contains the following metal oxides: SiO2, B2O3, alkali metal oxide e.g. K2O or/and Na2O and optionally Al2O3 and an alkaline earth metal oxide e.g. CaO. The contents of these metal oxides are preferably as follows: 50 to 95 mol-% SiO2, 0.2 to 30 mol-% B2O3, 0 to 10 mol-% Al2O3, 0 to 20 mol-% alkaline earth metal oxide and 0.2 to 20 mol-% alkali metal oxide where the percentages are each based on the total weight of the glass phase.
- A glass phase which contains SiO2, B2O3, K2O, Al2O3 and CaO has proven to be particularly suitable for the isolation of RNA. A glass phase which contains SiO2, B2O3 and Na2O has proven to be particularly suitable for the isolation of DNA.
- In the process according to the invention the adsorption matrix is preferably added in an amount which corresponds to the minimum amount required to quantitatively bind the analyte present in the sample, in particular a nucleic acid, or the amount is somewhat larger, preferably at most 50% and particularly preferably at most 20% above this amount. The expected amount of nucleic acid in various type of samples can—if it is not already known—be determined in advance by common techniques e.g. phenol/chloroform extraction and subsequent measurement of the optical density.
- Step (a) of the process according to the invention comprises lysing the sample in a reaction vessel. This lysis is usually carried out by lysing the cells present in the sample under denaturing conditions e.g. by adding a protease and a denaturing buffer. Proteinase K, pronase, elastase or/and lysozyme are preferably used as the proteinase. The use of proteinase K is particularly preferred.
- The protease digestion is carried out in a denaturing buffer which contains a chaotropic compound e.g. urea or urea derivatives, preferably a chaotropic salt,—particularly preferably a guanidinium salt such as guanidinium hydrochloride (especially for the isolation of DNA) or guanidinium thiocyanate (especially for the isolation of RNA) or a perchlorate or iodide. Concentrations in the range of 1 to 3 mol/l are preferred for guanidinium salts.
- In contrast to the method described in WO 96/41811 for sample preparation, the solid adsorption matrix is only added after lysing the sample. This procedure results in a significantly lower unspecific binding of undesired sample components, e.g. proteins, to the adsorption matrix.
- According to step (c) the analyte is selectively bound to the adsorption matrix by incubation in the lysing buffer preferably under chaotropic conditions.
- Step (d) of the process according to the invention comprises the separation of non-bound sample components from the adsorption matrix. For this purpose the non-bound sample components are preferably removed from the reaction vessel. This can be achieved by adding and removing a wash buffer, optionally several times, which preferably contains a quantity of at least 50% (v/v) and particularly preferably of at least 60% (v/v) of a solvent that is miscible with water such as ethanol, propanol and acetone.
- Steps (c), (d) or/and (e) of the process according to the invention are preferably carried out while mixing continuously or at intervals (i.e. mixing phases alternate with phases in which the reaction vessel is at rest) without adding external means. This mixing is preferably carried out by rotating the reaction vessel around its longitudinal axis while reversing the direction of rotation several times. The mixing vessel is particularly preferably rotated exactly around its longitudinal axis and the change in the direction of rotation is carried out such that the meniscus deflection of the liquid remains below□ a predetermined cut-off value. Such mixing processes are described in WO 91/15768 and EP-A-0 435 481.
- The duration of steps (c) or/and (e) is preferably 20 min at most and comprises a continuous mixing or an interval mixing in short cycles, preferably in short cycles of preferably two minutes maximum. Particularly good results were obtained by interval mixing in a one minute cycle comprising 20 sec mixing and 40 sec resting.
- When magnetic particles are used as an adsorption matrix it is possible to add liquids to the reaction vessel or aspirate liquids from the reaction vessel while mixing continuously, and the particles are held in the reaction vessel during the aspiration process. This mixing procedure allows the process according to the invention to be adjusted flexibly to suit various types of sample.
- Furthermore it ensures that there is always a homogeneous distribution of the magnetic particles in the liquid phase.
- Step (e) of the process according to the invention comprises the elution of the analyte from the adsorption matrix. A low salt buffer that is essentially free of organic solvents can be used for this as is known from the prior art. However, it was surprisingly found that the elution buffer can contain additional reagents such as enzymes e.g. enzymes used to manipulate nucleic acids such as RNases, DNases, restriction endonucleases, ligases, terminal transferases or/and polymerases. If the analyte is for example a DNA it is possible to add a DNase-free RNase during the elution in order to reduce the content of undesired RNA. On the other hand if the analyte is RNA, it is possible to add an RNase-free DNase during the elution. Other enzymes such as restriction endonucleases etc. can be added in an analogous manner. If the nucleic acid isolated by the process according to the invention is subjected to a subsequent amplification, a nucleic acid amplification master mix which contains the amplification buffer, nucleotides, primers, polymerase and buffer salts can also be added during the elution.
- Step (f) of the process according to the invention comprises separating the eluate from the adsorption matrix. This separation can be carried out in the usual manner e.g. by sedimentation but preferably by magnetic separation.
- The analytes isolated by the process according to the invention can be subsequently processed further in a known manner e.g. in the case of nucleic acids by amplification and subsequent detection, or detection without previous amplification or sequencing. For this purpose various analytes can be determined in aliquots of the eluate e.g. various viruses such as HIV, HCV and HBV.
- An important feature of the process according to the invention is that many or optionally even all steps can be carried out at essentially the same temperature i.e. within a temperature range of ±2.5° C. This temperature is preferably in the range of room temperature to 70° C., particular preferably from room temperature to 40° C., most preferably at room temperature i.e. ca. 18 to 32° C. In a preferred embodiment of the process according to the invention at least the steps (c) of adsorption and (d) of washing are carried out at this temperature. Other steps, in particular the steps (a) of lysing or/and (e) of elution are particularly preferably also carried out at this temperature. The entire sample preparation can for example be carried out at a uniform temperature for the determination of HIV in blood samples. Optionally an additional after-treatment step at an elevated temperature can take place after step (f) of the process according to the invention which improves the amplification yields for certain analytes. It may be necessary for other analytes to carry out the pre-treatment or/and the elution at an elevated temperature. In this case the elevated temperature is preferably in the range of more than 40° C. to 95° C. e.g. ca. 70° C.
- The process according to the invention is preferably carried out in an automated device. Examples of such devices are described in the following. It is also preferable that in the process according to the invention for sample preparation at least steps (a) to (e) are carried out in a single reaction vessel i.e. that there is no transfer into another reaction vessel. This considerably simplifies the process and also leads to a reduction of the risk of contamination.
- Yet a further subject matter of the invention is a reagent kit which is especially suitable for carrying out the process described above comprising
- (a) a protease,
- (b) a sample lysing buffer,
- (c) a wash buffer,
- (d) an elution buffer and
- (e) a suspension of magnetic glass particles.
- Yet an additional subject matter of the invention is a reagent kit for isolating DNA comprising magnetic glass particles whose glass phase contains SiO2, B2O3 and Na2O and a reagent kit for isolating RNA comprising magnetic glass particles whose glass phase contains SiO2, B2O3, Al2O3, CaO and Ka2O.
- Finally another subject matter of the present invention is a device for isolating an analyte from a biological sample comprising:
- a sample preparation device (1),
- a holding device for reagents (2),
- a first holding device for reaction vessels for sample preparation (3) which is equipped for an operating temperature of ≦70° C., in particular ≦40° C.,
- a second holding device for reaction vessels (4 a, 4 b, 4 c), which optionally contains a cooling or/and heating means,
- and a robotic tool device (5).
- The device according to the invention is preferably designed such that a single reaction vessel is used to carry out the 4 main steps of sample preparation i.e. lysis of a sample, adsorption of the released analyte e.g. a nucleic acid to a solid adsorption matrix e.g. magnetic glass particles, washing the adsorption matrix and eluting the analyte from the adsorption matrix.
- The device is designed such that the first holding device for holding the reaction vessels for sample preparation is used at least for the adsorption of the analyte to the solid adsorption matrix and for washing the adsorption matrix. In a preferred embodiment the first holding device is also used for the sample lysis or/and for the elution of the analyte from the adsorption matrix. The reaction vessels for the sample preparation have a volume of preferably at least 1 ml e.g. 1-5 ml.
- The second holding device is designed for reaction vessels to store or/and further process the analyte e.g. PCR vessels which usually have a different shape than the reaction vessels used for the sample preparation. The reaction vessels for storing or/and additional processing have a volume of preferably up to 500 μl, e.g. 50-200 μl. Furthermore the second holding device can contain vessels for reagents which are required to process the sample containing the analyte e.g. a PCR master mix.
- The device according to the invention can be designed such that one or several steps of the sample preparation or/and an after-treatment step can be carried out at an elevated temperature in the second holding device. For this purpose the second holding device can be designed to hold reaction vessels for at least one treatment step which is selected from lysing the sample, eluting the sample from the adsorption matrix and an after-treatment step after elution.
- The first holding device preferably contains means for the magnetic separation. In addition it is preferable that the first holding device contains means for mixing the reaction vessels in particular by rotating them around their longitudinal axis. Such means can optionally be provided for the second holding device.
- The robotic tool generally comprises automatic pipetting devices and optionally means for transporting reaction vessels e.g. between the first and second holding device. In addition a cap opening and closing unit may be integrated.
- Special embodiments of inventive devices are shown in detail in the following. In the embodiment shown in FIG.1 the sample preparation device (1) contains a holding device for reagents (2), a holding device for reaction vessels for sample preparation (3) with the functions mixing and magnetic separation which provides a temperature of preferably ≦40° C. and particularly preferably room temperature. The device additionally contains a holding station for further reaction vessels (4 a) e.g. for PCR vessels which has a temperature of 4° C. to room temperature. The device additionally contains automated devices for pipetting and handling reaction vessels (5) which enables movements in an X, Y and Z direction. In this embodiment of the device according to the invention the four main steps of sample preparation i.e. lysis, adsorption, washing and elution take place in a single reaction vessel in the first holding device. Eluates are stored and further reagents e.g. PCR master mix are added in the second holding device. For further processing e.g. for a subsequent PCR, the vessels are transferred to an appropriate device e.g. a thermocycler (not shown).
- In the embodiment shown in FIG. 2 the device contains a second holding device (4 b) which is designed to hold reaction vessels for further processing e.g. PCR vessels and is equipped to set a temperature of 4° C. (cooling the PCR master mix) to 95° C. to heat the eluate after elution from the adsorption matrix. A cap counter-heating is preferred to prevent condensation on the cap of the PCR vessels.
- The embodiment of the device according to the invention shown in FIG. 3 is provided with a second holding device for reaction vessels (4 c) which is designed to hold PCR vessels and sample preparation vessels. In this second holding device cooling e.g. to 40° C. and heating e.g. to 95° C. is possible to heat the lysate or/and the eluate. Also in this case a cap counter-heating is provided to prevent condensation on the cap of reaction vessels.
- In a further embodiment of the present invention (not shown) the first holding device is designed to set a temperature in the range of ≦70° C. The second holding device is—as shown in FIG. 3—suitable for cooling and heating sample processing and sample preparation vessels.
- The devices according to the invention can be used especially in a process as described above.
- The present application is additionally elucidated in more detail by the figures and examples.
- FIG. 1 shows a schematic representation of a first embodiment of the device according to the invention,
- FIG. 2 shows a schematic representation of a second embodiment of the device according to the invention,
- FIG. 3 shows a schematic representation of a third embodiment of the device according to the invention,
- FIG. 4 shows the result of a chlamydia detection by PCR using manual and semiautomatic sample preparation,
- FIG. 5 shows the result of a chlamydia detection by PCR using semiautomatic sample preparation and various temperature profiles during the sample preparation,
- FIG. 6 shows the result of a HIV detection by PCR using manual sample preparation (standard protocol) and semiautomated sample preparation at room temperature.
- 1. Preparation of Magnetic Glass Particles
- Two different sols were used. The sols were prepared as follows:
- Sol 1: (SiO2:B2O3:Na2O=40:8:2)
- Alcoholates of the oxides were stirred together in the above molar ratios analogously to the procedure in examples 1 and 2 of WO96/41811 to form a homogeneous phase. However, a deviation was that no HCl was used.
- Subsequently 30 g iriodin 600 Black Mica (Merk) in 100 ml sol was stirred in.
- Sol 2: (SiO2:B2O3:K2O:Al2O3:CaO=76:15:5:2:2)
- Alcoholates of the oxides were stirred together in the above molar ratios analogously to the procedure in examples 1 and 2 of WO96/41811 to form a homogeneous phase. However, a deviation was that no HCl was used.
- Subsequently 30 g iriodin 600 Black Mica (Merk) in 100 ml sol was stirred in.
- The sols were subsequently subjected to a spray drying process.
- The powder obtained by the spray drying was subjected to a separation of fines by sedimentation, a temperature treatment under a nitrogen atmosphere (60 l/h volume flow rate) at a heating rate of 1 K/min and kept for one hour at a compaction temperature in the range of 600 to 700° C. Subsequently the oven was cooled to 300° C. and flushed with oxygen for 1 h at this temperature. After cooling to room temperature the magnetic glass particles were removed and sieved through a 50 μm sieve to separate the coarse material.
- The magnetic glass particles obtained from
sol 1 are particularly suitable for the isolation of DNA. The glass particles obtained fromsol 2 are particularly suitable for the isolation of RNA. - 2. Standard Protocol for Sample Preparation for the Isolation of Nucleic Acids e.g. DNA
- The following standard protocol is suitable for isolating nucleic acids from biological samples such as whole blood or cultured cells. The nucleic acids obtained in this manner can be used directly after the elution for an amplification by PCR, a restriction cleavage or a Southern blot.
- The reaction kit contains:
- 1. binding buffer (4.7 mol/l guanidinium hydrochloride, 10 mmol/l urea, 10 mmol/l Tris HCl, 20% Triton®X-100, pH 5.7
- 2. lyophilized proteinase K (dissolved in H2O to a concentration of 20 mg/ml)
- 3. wash buffer (56% (v/v) ethanol, 20 mmol/l NaCl, 10 mmol/l Tris HCl, pH 7.5)
-
- 5. magnetic glass particles (MPG)
- a) tablets each containing 7.5 mg of the glass particles or
- b) 15% suspension of the glass particles in ethanol
- The kit components are stable and can be stored at room temperature. After dissolving proteinase K in water, the solution should be aliquoted and stored at −20° C. The frozen solution is stable for 12 months.
- Standard Protocol
- 1. 200 μl sample is added to a 2 ml reaction vessel and admixed with 200 μl binding buffer and 40 μl proteinase K solution. It is subsequently incubated for 10 min. The incubation is preferably carried out at room temperature. However, under certain circumstances the incubation temperature can also be increased to up to 70° C.
- 2. After the incubation 200 μl isopropanol and an MGP tablet (or alternatively 200 μl MGP suspension) is added and incubated for 5 min at room temperature.
- 3. The reaction vessel is placed in a magnetic particle separator (Boehringer Mannheim, Cat. No. 1 641 794) and separated for about 1 min.
- 4. The supernatant is discarded and the reaction vessels are removed from the MP separator.
- 5. After addition of 500 μl wash buffer, the contents of the reaction vessel are mixed and again placed in the MP separator for about 1 min.
- 6. The supernatant is discarded.
Step 5 is repeated three times. After the last washing process the remaining wash buffer is completely removed. - 7. For the
elution 100 μl of elution buffer which is optionally preheated to 70° C. is added. It is then mixed and incubated for 5 minutes at room temperature. The sample is placed in the MP separator and the supernatant is transferred into a clean reaction vessel. - 8. The nucleic acids e.g. DNA obtained in this manner are stable and can be subsequently directly processed further or stored at 4° C.
- The above protocol can also be used correspondingly for microtitre plates e.g. deep well microtitre plates (e.g. Ritter, J. J. Bioanalytic).
- 3. Chlamydia Trachomates DNA Detection by PCR
- 3.1 Manual Standard Protocol for Sample Preparation
- 200 μl of a urine sample and 240 μl binding buffer/proteinase K solution (5:1) are pipetted into a 2 ml reaction vessel, subjected to vortex mixing and incubated for 10 min at 70° C. Then the sample is cooled for 5 min to room temperature.
- 200 μl isopropanolic MGP solution is added to the sample by pipette. Immediately afterwards it is vortex mixed. The sample is then incubated for 15 min in a mixer e.g. Thermomixer 5436 (Eppendorf).
- The MGPs are concentrated by transferring the sample to a magnetic separator. After one minute the supernatant is completely removed by pipette.
- 0.5 ml wash buffer is added to the MGPs by pipette. The sample is subjected to vortex mixing and then transferred to the magnetic separator. The supernatant is removed by pipette after 1 min. The washing procedure is repeated for a further two times.
- 200 ml elution buffer is added to the MGP. The sample is incubated for 10 min at 70° C. in a thermomixer at 1400 rpm. Condensed water is collected by briefly centrifuging. The sample is transferred to the magnetic separator and after 1 min 180 μl eluate is removed. The eluate is pipetted into a new reaction vessel and stored at 4° C. (for a storage period of <24 h) or at −20° C. (for a longer storage period).
- 50 μl eluate is used for the PCR: The evaluation is by electrochemiluminescence.
- 3.2 Protocol for a Semiautomated Process
- Instead of the vortex mixing described in 3.1 and heating on a thermoblock, a semiautomated process is carried out in which the mixing and heating take place on a mixing and heating module. FIG. 4 shows a comparison of the determination of chlamydia (sample: 100 elementary antibodies per 100 ml urine; six-fold determination) between the manual standard protocol (vortex) and the semiautomated process (MTM). It can be seen that the sensitivity is not impaired by the automation.
- 3.3 Semiautomated Protocol at Room Temperature
- The sample preparation is carried out as described in section 3.2. However, the lysis and elution are carried out at room temperature.
- 3.4 Semiautomated Sample Preparation Protocol at Room Temperature with Subsequent After Treatment of the Eluate
- The sample preparation is carried out as described in section 3.3. After elution an incubation is carried out for 10 min at 70° C.
- FIG. 5 shows a comparison of the chlamydia determination (samples: SWE1, O chlamydia elementary antibodies (EAB) per ml urine, SWE2: 10 EAB, SWE3: 100 EAB and SWE 4:1000 EAB each per ml urine) between the sample preparation protocols described in sections 3.2, 3.3 and 3.4. It can be seen that the standard protocol is more sensitive compared to a sample preparation at room temperature (RT protocol MTM) for the determination of chlamydia. However, the results of the sample preparation at room temperature and subsequent aftertreatment of the eluate (RT protocol MTM with aftertreatment) show that this effect can be largely compensated. It is therefore surprising that a temperature step is not necessary during the sample preparation per se.
- This finding allows the sample preparation process to be considerably simplified since the steps of lysis, adsorption, washing and elution can be carried out at temperatures of ≦40° C. which simplifies an automation since a cap counter-heating and temperature regulation are not necessary.
- 4. HIV-RNA Detection by PCR
- 4.1 Manual Standard Protocol for Sample Preparation
- Frozen plasma is thawed for 5 min at 37° C. and cooled on ice for further processing.
- 50 μl of a proteinase K solution (25 mg/ml) is pipetted into a 1.5 ml Sarstedt reaction vessel. 250 μl sample is added to this and mixed in a vortex mixer. Then 300 μl lysis buffer is added and it is again vortex mixed.
- It is incubated for 10 min at room temperature on an Eppendorf mixer at 13,000 rpm. Then 300 μl of a MGP suspension (6 mg/ml MGP in isopropanol) is added, vortex mixed and incubated for 20 min at room temperature with continued mixing. The MGPs are separated on a magnetic separator and the supernatant is completely removed.
- 750 μl wash buffer is added to the MGPs. The MGPs are resuspended and separated as described previously. The wash procedure is repeated four times and the washing buffer is carefully removed at the end.
- Then 100 μ1 elution buffer is added and the MGPs are resuspended. After 15 minutes incubation at 80° C. on an Eppendorf thermomixer (13,000 rpm), 90 μl eluate is transferred into a new reaction vessel. 40 μl eluate is used for the subsequent HIV determination by RT-PCR.
- 4.2 Semiautomated Standard Protocol for Sample Preparation
- The sample preparation is carried out as described in section 4.1 except that the mixing and heating was carried out on a mixing and heating module.
- 4.3 Semiautomated Protocol at Room Temperature
- The sample preparation is essentially carried out as described in section 4.2 except that all steps are carried out at room temperature. The incubation period for lysis, adsorption and elution is in each case 15 min.
- It can be seen in FIG. 6 that the automation and the sample preparation at room temperature (RT protocol MTM) do not impair the sensitivity compared to the standard protocol with manual sample preparation (manual). Reproducible results are obtained for negative, low positive, moderately positive and highly positive plasma samples.
Claims (43)
1. Process for the isolation of an analyte from a biological sample comprising the steps:
(a) lysing the sample in a reaction vessel,
(b) adding a solid adsorption matrix
(c) incubating under such conditions that the analyte binds to the adsorption matrix,
(d) removing non-bound sample components from the reaction vessel,
(e) incubating under such conditions that the analyte is eluted from the adsorption matrix and
(f) separating the eluate from the adsorption matrix.
2. Process as claimed in claim 1 ,
wherein
step (a) comprises adding a protease and a denaturing buffer.
3. Process as claimed in claim 2 ,
wherein
proteinase K is used as the protease.
4. Process as claimed in claim 2 ,
wherein
a denaturing buffer is used which contains a guanidinium salt, in particular guanidinium hydrochloride or/and guanidinium thiocyanate.
5. Process as claimed in one of the previous claims,
wherein
magnetic glass particles are used as the solid adsorption matrix.
6. Process as claimed in claim 5 ,
wherein
the magnetic glass particles are added in the form of a suspension.
7. Process as claimed in claim 5 or 6,
wherein
glass particles are used whose glass phase contains SiO2, B2O3 and Na2O or SiO2, B2O3, Al2O3, CaO and K2O.
8. Process as claimed in one of the previous claims,
wherein
the amount of adsorption matrix added is at most 50% more than the amount that is required to quantitatively bind the analyte present in the sample.
9. Process as claimed in one of the previous claims,
wherein
a continuous or intermittent mixing without adding external devices is carried out at least during steps (c), (d) or/and (e).
10. Process as claimed in claim 9 ,
wherein
the mixing is achieved by rotating the reaction vessel around its longitudinal axis.
11. Process as claimed in claims 9 or 10,
wherein
the maximum period for carrying out steps (c) or/and (e) is 20 min in each case.
12. Process as claimed in one of the previous claims,
wherein
step (d) comprises adding and aspirating a wash buffer which is optionally repeated several times.
13. Process as claimed in claim 12 ,
wherein
a wash buffer is used with a content of at least 50% (v/v) of an organic solvent that is miscible with water.
14. Process as claimed in one of the previous claims,
wherein
additional reagents such as enzymes are added in step (e).
15. Process as claimed in one of the previous claims,
wherein
a low salt buffer is used for elution in step (e).
16. Process as claimed in one of the claims 1 to 14 ,
wherein
a nucleic acid amplification master mix is added for elution in step (e).
17. Process as claimed in one of the previous claims,
wherein
at least steps (c) and (d) are carried out at essentially the same temperature.
18. Process as claimed in claim 17 ,
wherein
step (a) or/and step (e) are also carried out at essentially the same temperature.
19. Process as claimed in claims 17 or 18,
wherein
the temperature is in the range from room temperature to 40° C.
20. Process as claimed in one of the claims 17 to 19 ,
wherein
the temperature is in the range of 18° C. to 32° C.
21. Process as claimed in one of the previous claims,
wherein
step (a) or/and step (e) are carried out at an elevated temperature.
22. Process as claimed in one of the previous claims,
wherein
an aftertreatment step at an elevated temperature is carried out after step (f).
23. Process as claimed in claims 21 or 22,
wherein
the elevated temperature is in the range from more than 40° C. to 95° C.
24. Process for the isolation of an analyte from a biological sample comprising the steps:
(a) lysing the sample in a reaction vessel,
(b) adding a solid adsorption matrix
(c) incubating under such conditions that the analyte binds to the adsorption matrix,
(d) separating non-bound sample components from the adsorption matrix,
(e) incubating under such conditions that the analyte is eluted from the adsorption matrix and
(f) separating the eluate from the adsorption matrix
wherein at least steps (c) and (d) are carried out at essentially the same temperature.
25. Process as claimed in one of the previous claims,
wherein
the analyte is a nucleic acid.
26. Process as claimed in one of the previous claims,
wherein
the process is carried out in an automated device.
27. Process as claimed in claims 1 or 24,
wherein
steps (a) to (e) are carried out in a single reaction vessel.
28. Reagent kit in particular to carry out the process as claimed in one of the claims 1 to 27 , comprising
(a) a protease,
(b) a sample lysing buffer,
(c) a wash buffer,
(d) an elution buffer and
(e) a suspension of magnetic glass particles.
29. Reagent kit for the isolation of DNA comprising magnetic glass particles whose glass phase contains SiO2, B2O3 and Na2O.
30. Reagent kit for the isolation of RNA comprising magnetic glass particles whose glass phase contains SiO2, B2O3, Al2O3, CaO and K2O.
31. Device for isolating an analyte from a biological sample comprising:
a sample preparation device (1),
a holding device for reagents (2),
a first holding device for reaction vessels for sample preparation (3) which is equipped for an operating temperature of ≦70° C., in particular ≦40° C.,
a second holding device for reaction vessels (4 a, 4 b, 4 c), which optionally contains a cooling or/and heating means,
and a robotic tool device (5).
32. Device as claimed in claim 31 ,
wherein
a single reaction vessel is used to lyse a sample, to adsorb the analyte to a solid adsorption matrix, to wash the adsorption matrix and to elute the analyte from the adsorption matrix.
33. Device as claimed in one of the claims 31 or 32,
wherein
the first holding device is used to hold the reaction vessels for the sample preparation and at least for the adsorption of the analyte to a solid adsorption matrix and for washing the adsorption matrix.
34. Device as claimed in claim 33 ,
wherein
the first holding device is addition ally used to hold the reaction vessels for sample preparation, for lysing the sample or/and for eluting the analyte from the adsorption matrix.
35. Device as claimed in one of the claims 31 to 34 ,
wherein
the second holding device for holding reaction vessels is used to store or/and further process the analyte.
36. Device as claimed in one of the claims 1 to 35 ,
wherein
the second holding device for holding vessels for reagents is used to process the sample further.
37. Device as claimed in one of the claims 31 to 36 ,
wherein
the second holding device for holding reaction vessels is used for at least one treatment step at an elevated temperature which is selected from lysing the sample, eluting the sample from the adsorption matrix and an aftertreatment step after elution.
38. Device as claimed in one of the claims 31 to 37 ,
wherein
the first holding device contains means for magnetic separation.
39. Device as claimed in one of the claims 31 to 38 ,
wherein
the first holding device contains means for mixing the reaction vessels by rotation around their longitudinal axis.
40. Device as claimed in one of the claims 31 to 39 ,
wherein
the robotic tool comprises automatic pipetting devices and optionally means for opening and closing reaction vessels.
41. Device as claimed in one of the claims 31 to 40 ,
wherein
the robotic tool comprises means for transporting reaction vessels between the first and second holding device.
42. Magnetic glass particles comprising a magnetic core and a glass coat which contains SiO2, B2O3, an alkali metal oxide and optionally Al2O3 and an alkaline earth metal oxide.
43. Glass particles as claimed in claim 41 ,
wherein
the glass coat contains SiO2, B2O3 and Na2O or SiO2, B2O3, Al2O3, K2O and CaO.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US10/426,641 US20030199078A1 (en) | 1997-10-01 | 2003-04-30 | Method, kit and apparatus for the isolation of nucleic acids |
Applications Claiming Priority (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
DE19743518A DE19743518A1 (en) | 1997-10-01 | 1997-10-01 | Automated, universally applicable sample preparation method |
DE19743518.1 | 1997-10-01 | ||
US09/509,750 US6562568B1 (en) | 1997-10-01 | 1998-09-29 | Method, kit and apparatus comprising magnetic glass particles for the isolation of biomolecules |
US10/426,641 US20030199078A1 (en) | 1997-10-01 | 2003-04-30 | Method, kit and apparatus for the isolation of nucleic acids |
Related Parent Applications (2)
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PCT/EP1998/006196 Continuation WO1999016781A2 (en) | 1997-10-01 | 1998-09-29 | Automatable method for preparing samples which can be universally applied |
US09/509,750 Continuation US6562568B1 (en) | 1997-10-01 | 1998-09-29 | Method, kit and apparatus comprising magnetic glass particles for the isolation of biomolecules |
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US20030199078A1 true US20030199078A1 (en) | 2003-10-23 |
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US10/426,641 Abandoned US20030199078A1 (en) | 1997-10-01 | 2003-04-30 | Method, kit and apparatus for the isolation of nucleic acids |
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US09/509,750 Expired - Lifetime US6562568B1 (en) | 1997-10-01 | 1998-09-29 | Method, kit and apparatus comprising magnetic glass particles for the isolation of biomolecules |
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US (2) | US6562568B1 (en) |
EP (2) | EP1783135B1 (en) |
JP (3) | JP4048022B2 (en) |
KR (1) | KR20010024366A (en) |
AT (1) | ATE366739T1 (en) |
AU (1) | AU1028299A (en) |
CA (1) | CA2305171C (en) |
DE (2) | DE19743518A1 (en) |
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ES (2) | ES2290996T3 (en) |
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US20050266462A1 (en) * | 1995-06-08 | 2005-12-01 | Roche Diagnostics Gmbh | Magnetic glass particles, method for their preparation and uses thereof |
US20070117972A1 (en) * | 1991-10-07 | 2007-05-24 | Invitrogen Corporation | Materials and Methods For the Purification of Polyelectrolytes, Particularly Nucleic Acids |
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Families Citing this family (59)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR100463475B1 (en) * | 1995-06-08 | 2005-06-22 | 로셰 디아그노스틱스 게엠베하 | Magnetic Pigment |
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US20050042660A1 (en) * | 2003-07-31 | 2005-02-24 | Hall Gerald Edward | Devices and methods for isolating RNA |
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US9175332B2 (en) | 2010-07-29 | 2015-11-03 | Roche Molecular Systems, Inc. | Generic PCR |
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CN107866286A (en) | 2013-03-15 | 2018-04-03 | 金马克诊断股份有限公司 | For manipulating system, the method and apparatus of deformable fluid container |
US9498778B2 (en) | 2014-11-11 | 2016-11-22 | Genmark Diagnostics, Inc. | Instrument for processing cartridge for performing assays in a closed sample preparation and reaction system |
USD881409S1 (en) | 2013-10-24 | 2020-04-14 | Genmark Diagnostics, Inc. | Biochip cartridge |
JP5928435B2 (en) | 2013-11-01 | 2016-06-01 | 株式会社安川電機 | Robot system, inspection method, and inspection object production method |
US10005080B2 (en) | 2014-11-11 | 2018-06-26 | Genmark Diagnostics, Inc. | Instrument and cartridge for performing assays in a closed sample preparation and reaction system employing electrowetting fluid manipulation |
US9598722B2 (en) | 2014-11-11 | 2017-03-21 | Genmark Diagnostics, Inc. | Cartridge for performing assays in a closed sample preparation and reaction system |
EP3218480A1 (en) | 2014-11-14 | 2017-09-20 | Corning Incorporated | Methods and kits for post-ivt rna purification |
EP3434372B1 (en) * | 2014-11-19 | 2020-02-26 | F. Hoffmann-La Roche AG | Particle mixing |
CN114381368B (en) * | 2021-12-31 | 2023-10-27 | 北京理工大学 | System and method for automatically separating biological analytes by utilizing gas-liquid interface |
Citations (92)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US2885366A (en) * | 1956-06-28 | 1959-05-05 | Du Pont | Product comprising a skin of dense, hydrated amorphous silica bound upon a core of another solid material and process of making same |
US2913419A (en) * | 1956-04-18 | 1959-11-17 | Du Pont | Chemical process and composition |
US3926659A (en) * | 1973-03-17 | 1975-12-16 | Merck Patent Gmbh | Iron-containing mica flake pigments |
US3945862A (en) * | 1973-06-26 | 1976-03-23 | Merck & Co., Inc. | Coated ferrous substrates comprising an amorphous magnesia-silica complex |
US4082905A (en) * | 1976-06-04 | 1978-04-04 | Bayer Aktiengesellschaft | Production of iron oxide pigments with improved resistance to oxidation |
US4124735A (en) * | 1976-12-02 | 1978-11-07 | Xerox Corporation | Magnetic glass carrier materials |
US4124385A (en) * | 1976-12-02 | 1978-11-07 | Xerox Corporation | Magnetic glass carrier materials |
US4126437A (en) * | 1976-12-02 | 1978-11-21 | Xerox Corporation | Magnetic glass carrier materials |
US4133169A (en) * | 1974-08-30 | 1979-01-09 | Ebauches S.A. | Electronic circuit for a quartz crystal watch |
US4280918A (en) * | 1980-03-10 | 1981-07-28 | International Business Machines Corporation | Magnetic particle dispersions |
US4297337A (en) * | 1979-04-13 | 1981-10-27 | Corning Glass Works | Solid-phase immunoassays using magnetic glass |
US4309459A (en) * | 1979-11-28 | 1982-01-05 | Tdk Electronics Co., Ltd. | Process for producing SiO2 coated iron oxide powder for use in the preparation of acicular magnetic iron or iron oxide powder |
US4336310A (en) * | 1980-01-28 | 1982-06-22 | Tdk Electronics Co., Ltd. | Magnetic recording medium and preparation thereof |
US4360441A (en) * | 1981-06-25 | 1982-11-23 | Corning Glass Works | Glass-encapsulated magnetic materials and methods for making them |
US4395271A (en) * | 1979-04-13 | 1983-07-26 | Corning Glass Works | Method for making porous magnetic glass and crystal-containing structures |
US4454234A (en) * | 1981-12-30 | 1984-06-12 | Czerlinski George H | Coated magnetizable microparticles, reversible suspensions thereof, and processes relating thereto |
US4477492A (en) * | 1983-04-22 | 1984-10-16 | E. I. Du Pont De Nemours And Company | Process for preparing superficially porous supports for chromatography and catalysts |
US4554088A (en) * | 1983-05-12 | 1985-11-19 | Advanced Magnetics Inc. | Magnetic particles for use in separations |
US4564537A (en) * | 1984-01-21 | 1986-01-14 | The British Petroleum Company P.L.C. | Process for depositing a silica coating on a metal surface |
US4628037A (en) * | 1983-05-12 | 1986-12-09 | Advanced Magnetics, Inc. | Binding assays employing magnetic particles |
US4672040A (en) * | 1983-05-12 | 1987-06-09 | Advanced Magnetics, Inc. | Magnetic particles for use in separations |
US4683202A (en) * | 1985-03-28 | 1987-07-28 | Cetus Corporation | Process for amplifying nucleic acid sequences |
US4683195A (en) * | 1986-01-30 | 1987-07-28 | Cetus Corporation | Process for amplifying, detecting, and/or-cloning nucleic acid sequences |
US4695392A (en) * | 1983-05-12 | 1987-09-22 | Advanced Magnetics Inc. | Magnetic particles for use in separations |
US4695393A (en) * | 1983-05-12 | 1987-09-22 | Advanced Magnetics Inc. | Magnetic particles for use in separations |
US4698302A (en) * | 1983-05-12 | 1987-10-06 | Advanced Magnetics, Inc. | Enzymatic reactions using magnetic particles |
US4699171A (en) * | 1986-12-19 | 1987-10-13 | Sundstrand Corporation | Multiple port relief valve |
US4748121A (en) * | 1984-11-30 | 1988-05-31 | Ppg Industries, Inc. | Porous glass fibers with immobilized biochemically active material |
US4751211A (en) * | 1986-08-07 | 1988-06-14 | Aluminum Company Of America | Composite adsorbent for removing acids from organophosphate functional fluids |
US4767670A (en) * | 1987-01-21 | 1988-08-30 | E. I. Du Pont De Nemours And Company | Chromatographic supports for separation of oligonucleotides |
US4804561A (en) * | 1986-10-25 | 1989-02-14 | Chisso Corporation | Process for producing ferromagnetic metal fine particles |
US4824712A (en) * | 1984-07-16 | 1989-04-25 | Ppg Industries, Inc. | Treatment of glass to reduce venting during thermal treatment and a glass article made thereby |
US4910148A (en) * | 1987-02-10 | 1990-03-20 | Dynal A. S. | Magnetic separation of magnetized particles from biological fluids |
US5039559A (en) * | 1988-05-24 | 1991-08-13 | Sang Jean V | Method of making magnetically attractable particles |
US5041390A (en) * | 1988-08-11 | 1991-08-20 | Skov Per S | Method and means for allergy diagnosis |
US5055194A (en) * | 1989-07-28 | 1991-10-08 | University Of Pennsylvania | Support for high performance liquid chromatography in a magnetically stabilized fluidized bed |
US5057426A (en) * | 1986-11-22 | 1991-10-15 | Diagen Institut Fur Molekular-Biologische, Diagnostik Gmbh | Method for separating long-chain nucleic acids |
US5075430A (en) * | 1988-12-12 | 1991-12-24 | Bio-Rad Laboratories, Inc. | Process for the purification of DNA on diatomaceous earth |
US5076950A (en) * | 1985-12-20 | 1991-12-31 | Syntex (U.S.A.) Inc. | Magnetic composition for particle separation |
US5155018A (en) * | 1991-07-10 | 1992-10-13 | Hahnemann University | Process and kit for isolating and purifying RNA from biological sources |
US5206568A (en) * | 1986-03-26 | 1993-04-27 | Beckman Instruments, Inc. | Coordinated control of stepper motors |
US5210015A (en) * | 1990-08-06 | 1993-05-11 | Hoffman-La Roche Inc. | Homogeneous assay system using the nuclease activity of a nucleic acid polymerase |
US5217804A (en) * | 1990-11-06 | 1993-06-08 | Eastman Kodak Company | Magnetic particles |
US5234809A (en) * | 1989-03-23 | 1993-08-10 | Akzo N.V. | Process for isolating nucleic acid |
US5236623A (en) * | 1984-07-11 | 1993-08-17 | Rhone-Poulenc Chimie | Process for the production of a silica colloid |
US5279936A (en) * | 1989-12-22 | 1994-01-18 | Syntex (U.S.A.) Inc. | Method of separation employing magnetic particles and second medium |
US5312485A (en) * | 1988-08-05 | 1994-05-17 | J. M. Huber Corporation | Precipitated encapsulated paper pigments and methods |
US5316699A (en) * | 1990-03-28 | 1994-05-31 | The United States Of America As Repesented By The Secretary Of Commerce | Process for the controlled preparation of a composite of ultrafine magnetic particles homogeneously dispersed in a dielectric matrix |
US5340393A (en) * | 1992-04-28 | 1994-08-23 | E. I. Du Pont De Nemours And Company | Process for preparing silica coated inorganic particles |
US5346994A (en) * | 1992-01-28 | 1994-09-13 | Piotr Chomczynski | Shelf-stable product and process for isolating RNA, DNA and proteins |
US5352645A (en) * | 1989-04-14 | 1994-10-04 | E. I. Du Pont De Nemours And Company | Silica microspheres, method of improving attrition resistance and use |
US5368933A (en) * | 1990-02-21 | 1994-11-29 | Toda Kogyo Corp. | Superparamagnetic fine particles of iron oxide and magnetic recording media containing said particles |
US5389482A (en) * | 1992-04-23 | 1995-02-14 | Toda Kogyo Corp. | Magnetic particle powder included in magnetic toners for magnetic image character recognition |
US5395498A (en) * | 1991-11-06 | 1995-03-07 | Gombinsky; Moshe | Method for separating biological macromolecules and means therfor |
US5438127A (en) * | 1993-09-27 | 1995-08-01 | Becton Dickinson And Company | DNA purification by solid phase extraction using a PCl3 modified glass fiber membrane |
US5443791A (en) * | 1990-04-06 | 1995-08-22 | Perkin Elmer - Applied Biosystems Division | Automated molecular biology laboratory |
US5458813A (en) * | 1992-07-28 | 1995-10-17 | Enichem S.P.A. | Method for preparing boron-containing porous gels |
US5470660A (en) * | 1992-10-06 | 1995-11-28 | Toda Kogyo Corporation | Iron oxide particles and process for producing the same |
US5503816A (en) * | 1993-09-27 | 1996-04-02 | Becton Dickinson And Company | Silicate compounds for DNA purification |
US5512332A (en) * | 1985-10-04 | 1996-04-30 | Immunivest Corporation | Process of making resuspendable coated magnetic particles |
US5520899A (en) * | 1992-11-13 | 1996-05-28 | Becton, Dickinson And Company | Process for preparing boron, aluminum, and phosphorus silicates for purification of DNA |
US5578238A (en) * | 1992-10-30 | 1996-11-26 | Lord Corporation | Magnetorheological materials utilizing surface-modified particles |
US5597531A (en) * | 1985-10-04 | 1997-01-28 | Immunivest Corporation | Resuspendable coated magnetic particles and stable magnetic particle suspensions |
US5599627A (en) * | 1993-10-08 | 1997-02-04 | Toda Kogyo Corporation | Magnetic particles comprising magnetite core and process for producing the same |
US5610274A (en) * | 1991-11-20 | 1997-03-11 | Cpg, Inc. | Production and use of magnetic porous inorganic materials |
US5648170A (en) * | 1993-04-27 | 1997-07-15 | Toda Kogyo Corporation | Coated granular magnetite particles and process for producing the same |
US5658548A (en) * | 1993-08-30 | 1997-08-19 | Promega Corporation | Nucleic acid purification on silica gel and glass mixtures |
US5660984A (en) * | 1994-12-09 | 1997-08-26 | Davis; Thomas E. | DNA isolating apparatus comprising a non-porous DNA binding, anion exchange resin and methods of use thereof |
US5665554A (en) * | 1994-06-09 | 1997-09-09 | Amersham International Plc | Magnetic bead precipitation method |
US5681946A (en) * | 1990-02-13 | 1997-10-28 | Amersham International Plc | Precipitating polymers |
US5683875A (en) * | 1995-05-04 | 1997-11-04 | Hewlett-Packard Company | Method for detecting a target nucleic acid analyte in a sample |
US5705137A (en) * | 1995-06-01 | 1998-01-06 | Degussa Aktiengesellschaft | Precipitated silicas, a process for their preparation and their use in vulcanizable rubber mixtures |
US5723591A (en) * | 1994-11-16 | 1998-03-03 | Perkin-Elmer Corporation | Self-quenching fluorescence probe |
US5734020A (en) * | 1991-11-20 | 1998-03-31 | Cpg, Inc. | Production and use of magnetic porous inorganic materials |
US5747663A (en) * | 1994-02-07 | 1998-05-05 | Qiagen Gmbh | Process for the depletion or removal of endotoxins |
US5763173A (en) * | 1990-06-11 | 1998-06-09 | Nexstar Pharmaceuticals, Inc. | Nucleic acid ligand inhibitors to DNA polymerases |
US5783686A (en) * | 1995-09-15 | 1998-07-21 | Beckman Instruments, Inc. | Method for purifying nucleic acids from heterogenous mixtures |
US5898071A (en) * | 1994-09-20 | 1999-04-27 | Whitehead Institute For Biomedical Research | DNA purification and isolation using magnetic particles |
US5904848A (en) * | 1996-02-21 | 1999-05-18 | Cpg, Inc. | Controlled pore glass-synthetic resin membrane |
US5925573A (en) * | 1995-03-21 | 1999-07-20 | Bio Merieux | Method and device for the determination of an analyte using super paramagnetic reactive particles |
US5928958A (en) * | 1994-07-27 | 1999-07-27 | Pilgrimm; Herbert | Superparamagnetic particles, process for their manufacture and usage |
US5945525A (en) * | 1995-07-07 | 1999-08-31 | Toyo Boseki Kabushiki Kaisha | Method for isolating nucleic acids using silica-coated magnetic particles |
US5972721A (en) * | 1996-03-14 | 1999-10-26 | The United States Of America As Represented By The Secretary Of The Air Force | Immunomagnetic assay system for clinical diagnosis and other purposes |
US5990301A (en) * | 1994-02-07 | 1999-11-23 | Qiagen Gmbh | Process for the separation and purification of nucleic acids from biological sources |
US5990479A (en) * | 1997-11-25 | 1999-11-23 | Regents Of The University Of California | Organo Luminescent semiconductor nanocrystal probes for biological applications and process for making and using such probes |
US6027945A (en) * | 1997-01-21 | 2000-02-22 | Promega Corporation | Methods of isolating biological target materials using silica magnetic particles |
US6136083A (en) * | 1995-06-08 | 2000-10-24 | Institut Fuer Neue Materialien Gemeinnuetzige Gmbh | Coated inorganic pigments, process for their production and their use |
US6255477B1 (en) * | 1995-06-08 | 2001-07-03 | Roche Diagnostics Gmbh | Particles having a magnetic core and outer glass layer for separating biological material |
US6274386B1 (en) * | 1996-06-07 | 2001-08-14 | Roche Diagnostics Gmbh | Reagent preparation containing magnetic particles in tablet form |
US6296937B2 (en) * | 1997-01-21 | 2001-10-02 | W. R. Grace & Co.-Conn. | Silica adsorbent on magnetic substrate |
US6545143B1 (en) * | 1998-11-30 | 2003-04-08 | Roche Diagnostics, Gmbh | Magnetic particles for purifying nucleic acids |
US20030096984A1 (en) * | 2000-03-31 | 2003-05-22 | Cully Doris F. | Dna molecules encoding ligand gated ion channels from dermacentor variabilis |
Family Cites Families (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4448884A (en) * | 1982-03-03 | 1984-05-15 | Kms Fusion, Inc. | Glass-surface microcarrier for growth of cell cultures |
CH672445A5 (en) | 1987-02-27 | 1989-11-30 | Hratch Boyadjian | |
KR960000479B1 (en) * | 1987-03-02 | 1996-01-08 | 젠-프로우브 인코오퍼레이티드 | Polycationic supports for nucleic acid purification separation and hybridization |
CA2231861A1 (en) | 1995-09-13 | 1997-03-20 | Takanori Oka | Method for concentrating mutant nucleic acid and nucleic acid-concentrating assay kit for said concentration method |
US6242235B1 (en) | 1998-06-24 | 2001-06-05 | Promega Corp. | Polymerase stabilization by polyethoxylated amine surfactants |
-
1997
- 1997-10-01 DE DE19743518A patent/DE19743518A1/en not_active Ceased
-
1998
- 1998-09-29 EP EP07003878A patent/EP1783135B1/en not_active Expired - Lifetime
- 1998-09-29 EP EP98952670A patent/EP1019430B1/en not_active Expired - Lifetime
- 1998-09-29 PT PT98952670T patent/PT1019430E/en unknown
- 1998-09-29 DK DK98952670T patent/DK1019430T3/en active
- 1998-09-29 AU AU10282/99A patent/AU1028299A/en not_active Abandoned
- 1998-09-29 ES ES98952670T patent/ES2290996T3/en not_active Expired - Lifetime
- 1998-09-29 CA CA002305171A patent/CA2305171C/en not_active Expired - Lifetime
- 1998-09-29 IL IL13529998A patent/IL135299A0/en unknown
- 1998-09-29 KR KR1020007003473A patent/KR20010024366A/en not_active Application Discontinuation
- 1998-09-29 JP JP2000513863A patent/JP4048022B2/en not_active Expired - Lifetime
- 1998-09-29 AT AT98952670T patent/ATE366739T1/en active
- 1998-09-29 US US09/509,750 patent/US6562568B1/en not_active Expired - Lifetime
- 1998-09-29 DE DE59814056T patent/DE59814056D1/en not_active Expired - Lifetime
- 1998-09-29 ES ES07003878T patent/ES2398713T3/en not_active Expired - Lifetime
- 1998-09-29 WO PCT/EP1998/006196 patent/WO1999016781A2/en not_active Application Discontinuation
-
2000
- 2000-03-27 IL IL135299A patent/IL135299A/en not_active IP Right Cessation
-
2003
- 2003-04-30 US US10/426,641 patent/US20030199078A1/en not_active Abandoned
-
2007
- 2007-02-27 JP JP2007047875A patent/JP2007175060A/en active Pending
- 2007-10-29 JP JP2007280179A patent/JP4659013B2/en not_active Expired - Lifetime
Patent Citations (102)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US2913419A (en) * | 1956-04-18 | 1959-11-17 | Du Pont | Chemical process and composition |
US2885366A (en) * | 1956-06-28 | 1959-05-05 | Du Pont | Product comprising a skin of dense, hydrated amorphous silica bound upon a core of another solid material and process of making same |
US3926659A (en) * | 1973-03-17 | 1975-12-16 | Merck Patent Gmbh | Iron-containing mica flake pigments |
US3945862A (en) * | 1973-06-26 | 1976-03-23 | Merck & Co., Inc. | Coated ferrous substrates comprising an amorphous magnesia-silica complex |
US4133169A (en) * | 1974-08-30 | 1979-01-09 | Ebauches S.A. | Electronic circuit for a quartz crystal watch |
US4082905A (en) * | 1976-06-04 | 1978-04-04 | Bayer Aktiengesellschaft | Production of iron oxide pigments with improved resistance to oxidation |
US4124735A (en) * | 1976-12-02 | 1978-11-07 | Xerox Corporation | Magnetic glass carrier materials |
US4124385A (en) * | 1976-12-02 | 1978-11-07 | Xerox Corporation | Magnetic glass carrier materials |
US4126437A (en) * | 1976-12-02 | 1978-11-21 | Xerox Corporation | Magnetic glass carrier materials |
US4297337A (en) * | 1979-04-13 | 1981-10-27 | Corning Glass Works | Solid-phase immunoassays using magnetic glass |
US4395271A (en) * | 1979-04-13 | 1983-07-26 | Corning Glass Works | Method for making porous magnetic glass and crystal-containing structures |
US4309459A (en) * | 1979-11-28 | 1982-01-05 | Tdk Electronics Co., Ltd. | Process for producing SiO2 coated iron oxide powder for use in the preparation of acicular magnetic iron or iron oxide powder |
US4336310A (en) * | 1980-01-28 | 1982-06-22 | Tdk Electronics Co., Ltd. | Magnetic recording medium and preparation thereof |
US4280918A (en) * | 1980-03-10 | 1981-07-28 | International Business Machines Corporation | Magnetic particle dispersions |
US4360441A (en) * | 1981-06-25 | 1982-11-23 | Corning Glass Works | Glass-encapsulated magnetic materials and methods for making them |
US4454234A (en) * | 1981-12-30 | 1984-06-12 | Czerlinski George H | Coated magnetizable microparticles, reversible suspensions thereof, and processes relating thereto |
US4477492A (en) * | 1983-04-22 | 1984-10-16 | E. I. Du Pont De Nemours And Company | Process for preparing superficially porous supports for chromatography and catalysts |
US4554088A (en) * | 1983-05-12 | 1985-11-19 | Advanced Magnetics Inc. | Magnetic particles for use in separations |
US4695392A (en) * | 1983-05-12 | 1987-09-22 | Advanced Magnetics Inc. | Magnetic particles for use in separations |
US4628037A (en) * | 1983-05-12 | 1986-12-09 | Advanced Magnetics, Inc. | Binding assays employing magnetic particles |
US4672040A (en) * | 1983-05-12 | 1987-06-09 | Advanced Magnetics, Inc. | Magnetic particles for use in separations |
US4698302A (en) * | 1983-05-12 | 1987-10-06 | Advanced Magnetics, Inc. | Enzymatic reactions using magnetic particles |
US4695393A (en) * | 1983-05-12 | 1987-09-22 | Advanced Magnetics Inc. | Magnetic particles for use in separations |
US4564537A (en) * | 1984-01-21 | 1986-01-14 | The British Petroleum Company P.L.C. | Process for depositing a silica coating on a metal surface |
US5236623A (en) * | 1984-07-11 | 1993-08-17 | Rhone-Poulenc Chimie | Process for the production of a silica colloid |
US4824712A (en) * | 1984-07-16 | 1989-04-25 | Ppg Industries, Inc. | Treatment of glass to reduce venting during thermal treatment and a glass article made thereby |
US4748121A (en) * | 1984-11-30 | 1988-05-31 | Ppg Industries, Inc. | Porous glass fibers with immobilized biochemically active material |
US4683202A (en) * | 1985-03-28 | 1987-07-28 | Cetus Corporation | Process for amplifying nucleic acid sequences |
US4683202B1 (en) * | 1985-03-28 | 1990-11-27 | Cetus Corp | |
US5512332A (en) * | 1985-10-04 | 1996-04-30 | Immunivest Corporation | Process of making resuspendable coated magnetic particles |
US5597531A (en) * | 1985-10-04 | 1997-01-28 | Immunivest Corporation | Resuspendable coated magnetic particles and stable magnetic particle suspensions |
US5076950A (en) * | 1985-12-20 | 1991-12-31 | Syntex (U.S.A.) Inc. | Magnetic composition for particle separation |
US4683195A (en) * | 1986-01-30 | 1987-07-28 | Cetus Corporation | Process for amplifying, detecting, and/or-cloning nucleic acid sequences |
US4683195B1 (en) * | 1986-01-30 | 1990-11-27 | Cetus Corp | |
US5206568A (en) * | 1986-03-26 | 1993-04-27 | Beckman Instruments, Inc. | Coordinated control of stepper motors |
US4751211A (en) * | 1986-08-07 | 1988-06-14 | Aluminum Company Of America | Composite adsorbent for removing acids from organophosphate functional fluids |
US4804561A (en) * | 1986-10-25 | 1989-02-14 | Chisso Corporation | Process for producing ferromagnetic metal fine particles |
US5057426A (en) * | 1986-11-22 | 1991-10-15 | Diagen Institut Fur Molekular-Biologische, Diagnostik Gmbh | Method for separating long-chain nucleic acids |
US4699171A (en) * | 1986-12-19 | 1987-10-13 | Sundstrand Corporation | Multiple port relief valve |
US4767670A (en) * | 1987-01-21 | 1988-08-30 | E. I. Du Pont De Nemours And Company | Chromatographic supports for separation of oligonucleotides |
US4910148A (en) * | 1987-02-10 | 1990-03-20 | Dynal A. S. | Magnetic separation of magnetized particles from biological fluids |
US5039559A (en) * | 1988-05-24 | 1991-08-13 | Sang Jean V | Method of making magnetically attractable particles |
US5662824A (en) * | 1988-05-24 | 1997-09-02 | Alfa Biotech Spa | Magnetically attractable particles and method |
US5312485A (en) * | 1988-08-05 | 1994-05-17 | J. M. Huber Corporation | Precipitated encapsulated paper pigments and methods |
US5041390A (en) * | 1988-08-11 | 1991-08-20 | Skov Per S | Method and means for allergy diagnosis |
US5075430A (en) * | 1988-12-12 | 1991-12-24 | Bio-Rad Laboratories, Inc. | Process for the purification of DNA on diatomaceous earth |
US5234809A (en) * | 1989-03-23 | 1993-08-10 | Akzo N.V. | Process for isolating nucleic acid |
US5352645A (en) * | 1989-04-14 | 1994-10-04 | E. I. Du Pont De Nemours And Company | Silica microspheres, method of improving attrition resistance and use |
US5055194A (en) * | 1989-07-28 | 1991-10-08 | University Of Pennsylvania | Support for high performance liquid chromatography in a magnetically stabilized fluidized bed |
US5279936A (en) * | 1989-12-22 | 1994-01-18 | Syntex (U.S.A.) Inc. | Method of separation employing magnetic particles and second medium |
US5681946A (en) * | 1990-02-13 | 1997-10-28 | Amersham International Plc | Precipitating polymers |
US5368933A (en) * | 1990-02-21 | 1994-11-29 | Toda Kogyo Corp. | Superparamagnetic fine particles of iron oxide and magnetic recording media containing said particles |
US5316699A (en) * | 1990-03-28 | 1994-05-31 | The United States Of America As Repesented By The Secretary Of Commerce | Process for the controlled preparation of a composite of ultrafine magnetic particles homogeneously dispersed in a dielectric matrix |
US5443791A (en) * | 1990-04-06 | 1995-08-22 | Perkin Elmer - Applied Biosystems Division | Automated molecular biology laboratory |
US5763173A (en) * | 1990-06-11 | 1998-06-09 | Nexstar Pharmaceuticals, Inc. | Nucleic acid ligand inhibitors to DNA polymerases |
US5210015A (en) * | 1990-08-06 | 1993-05-11 | Hoffman-La Roche Inc. | Homogeneous assay system using the nuclease activity of a nucleic acid polymerase |
US5804375A (en) * | 1990-08-06 | 1998-09-08 | Roche Molecular Systems, Inc. | Reaction mixtures for detection of target nucleic acids |
US5487972A (en) * | 1990-08-06 | 1996-01-30 | Hoffmann-La Roche Inc. | Nucleic acid detection by the 5'-3'exonuclease activity of polymerases acting on adjacently hybridized oligonucleotides |
US5217804A (en) * | 1990-11-06 | 1993-06-08 | Eastman Kodak Company | Magnetic particles |
US5155018A (en) * | 1991-07-10 | 1992-10-13 | Hahnemann University | Process and kit for isolating and purifying RNA from biological sources |
US5395498A (en) * | 1991-11-06 | 1995-03-07 | Gombinsky; Moshe | Method for separating biological macromolecules and means therfor |
US5734020A (en) * | 1991-11-20 | 1998-03-31 | Cpg, Inc. | Production and use of magnetic porous inorganic materials |
US5610274A (en) * | 1991-11-20 | 1997-03-11 | Cpg, Inc. | Production and use of magnetic porous inorganic materials |
US5346994A (en) * | 1992-01-28 | 1994-09-13 | Piotr Chomczynski | Shelf-stable product and process for isolating RNA, DNA and proteins |
US5389482A (en) * | 1992-04-23 | 1995-02-14 | Toda Kogyo Corp. | Magnetic particle powder included in magnetic toners for magnetic image character recognition |
US5340393A (en) * | 1992-04-28 | 1994-08-23 | E. I. Du Pont De Nemours And Company | Process for preparing silica coated inorganic particles |
US5458813A (en) * | 1992-07-28 | 1995-10-17 | Enichem S.P.A. | Method for preparing boron-containing porous gels |
US5470660A (en) * | 1992-10-06 | 1995-11-28 | Toda Kogyo Corporation | Iron oxide particles and process for producing the same |
US5512405A (en) * | 1992-10-06 | 1996-04-30 | Toda Kogyo Corporation | Iron oxide particles and process for producing the same |
US5578238A (en) * | 1992-10-30 | 1996-11-26 | Lord Corporation | Magnetorheological materials utilizing surface-modified particles |
US5520899A (en) * | 1992-11-13 | 1996-05-28 | Becton, Dickinson And Company | Process for preparing boron, aluminum, and phosphorus silicates for purification of DNA |
US5648170A (en) * | 1993-04-27 | 1997-07-15 | Toda Kogyo Corporation | Coated granular magnetite particles and process for producing the same |
US5658548A (en) * | 1993-08-30 | 1997-08-19 | Promega Corporation | Nucleic acid purification on silica gel and glass mixtures |
US5658548C1 (en) * | 1993-08-30 | 2001-07-24 | Promega Corp | Nucleic acid purification on silica geland glass mixtures |
US5503816A (en) * | 1993-09-27 | 1996-04-02 | Becton Dickinson And Company | Silicate compounds for DNA purification |
US5438127A (en) * | 1993-09-27 | 1995-08-01 | Becton Dickinson And Company | DNA purification by solid phase extraction using a PCl3 modified glass fiber membrane |
US5599627A (en) * | 1993-10-08 | 1997-02-04 | Toda Kogyo Corporation | Magnetic particles comprising magnetite core and process for producing the same |
US5747663A (en) * | 1994-02-07 | 1998-05-05 | Qiagen Gmbh | Process for the depletion or removal of endotoxins |
US5990301A (en) * | 1994-02-07 | 1999-11-23 | Qiagen Gmbh | Process for the separation and purification of nucleic acids from biological sources |
US5665554A (en) * | 1994-06-09 | 1997-09-09 | Amersham International Plc | Magnetic bead precipitation method |
US5928958A (en) * | 1994-07-27 | 1999-07-27 | Pilgrimm; Herbert | Superparamagnetic particles, process for their manufacture and usage |
US5898071A (en) * | 1994-09-20 | 1999-04-27 | Whitehead Institute For Biomedical Research | DNA purification and isolation using magnetic particles |
US5723591A (en) * | 1994-11-16 | 1998-03-03 | Perkin-Elmer Corporation | Self-quenching fluorescence probe |
US5660984A (en) * | 1994-12-09 | 1997-08-26 | Davis; Thomas E. | DNA isolating apparatus comprising a non-porous DNA binding, anion exchange resin and methods of use thereof |
US5925573A (en) * | 1995-03-21 | 1999-07-20 | Bio Merieux | Method and device for the determination of an analyte using super paramagnetic reactive particles |
US5683875A (en) * | 1995-05-04 | 1997-11-04 | Hewlett-Packard Company | Method for detecting a target nucleic acid analyte in a sample |
US5705137A (en) * | 1995-06-01 | 1998-01-06 | Degussa Aktiengesellschaft | Precipitated silicas, a process for their preparation and their use in vulcanizable rubber mixtures |
US6870047B2 (en) * | 1995-06-08 | 2005-03-22 | Roche Diagnostics Gmbh | Magnetic pigment |
US6136083A (en) * | 1995-06-08 | 2000-10-24 | Institut Fuer Neue Materialien Gemeinnuetzige Gmbh | Coated inorganic pigments, process for their production and their use |
US6255477B1 (en) * | 1995-06-08 | 2001-07-03 | Roche Diagnostics Gmbh | Particles having a magnetic core and outer glass layer for separating biological material |
US5945525A (en) * | 1995-07-07 | 1999-08-31 | Toyo Boseki Kabushiki Kaisha | Method for isolating nucleic acids using silica-coated magnetic particles |
US5783686A (en) * | 1995-09-15 | 1998-07-21 | Beckman Instruments, Inc. | Method for purifying nucleic acids from heterogenous mixtures |
US5904848A (en) * | 1996-02-21 | 1999-05-18 | Cpg, Inc. | Controlled pore glass-synthetic resin membrane |
US5972721A (en) * | 1996-03-14 | 1999-10-26 | The United States Of America As Represented By The Secretary Of The Air Force | Immunomagnetic assay system for clinical diagnosis and other purposes |
US6274386B1 (en) * | 1996-06-07 | 2001-08-14 | Roche Diagnostics Gmbh | Reagent preparation containing magnetic particles in tablet form |
US6027945A (en) * | 1997-01-21 | 2000-02-22 | Promega Corporation | Methods of isolating biological target materials using silica magnetic particles |
US6296937B2 (en) * | 1997-01-21 | 2001-10-02 | W. R. Grace & Co.-Conn. | Silica adsorbent on magnetic substrate |
US6368800B1 (en) * | 1997-01-21 | 2002-04-09 | Promega Corporation | Kits for isolating biological target materials using silica magnetic particles |
US5990479A (en) * | 1997-11-25 | 1999-11-23 | Regents Of The University Of California | Organo Luminescent semiconductor nanocrystal probes for biological applications and process for making and using such probes |
US6545143B1 (en) * | 1998-11-30 | 2003-04-08 | Roche Diagnostics, Gmbh | Magnetic particles for purifying nucleic acids |
US6919444B2 (en) * | 1998-11-30 | 2005-07-19 | Roche Diagnostics Gmbh | Magnetic particles for purifying nucleic acids |
US20030096984A1 (en) * | 2000-03-31 | 2003-05-22 | Cully Doris F. | Dna molecules encoding ligand gated ion channels from dermacentor variabilis |
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JP4659013B2 (en) | 2011-03-30 |
JP2007175060A (en) | 2007-07-12 |
DE59814056D1 (en) | 2007-08-23 |
ES2398713T3 (en) | 2013-03-21 |
CA2305171C (en) | 2008-06-17 |
EP1783135B1 (en) | 2012-11-07 |
PT1019430E (en) | 2007-10-16 |
EP1783135A1 (en) | 2007-05-09 |
ATE366739T1 (en) | 2007-08-15 |
IL135299A0 (en) | 2001-05-20 |
JP2008134239A (en) | 2008-06-12 |
US6562568B1 (en) | 2003-05-13 |
JP4048022B2 (en) | 2008-02-13 |
WO1999016781A2 (en) | 1999-04-08 |
WO1999016781A3 (en) | 1999-09-16 |
IL135299A (en) | 2010-11-30 |
ES2290996T3 (en) | 2008-02-16 |
EP1019430A2 (en) | 2000-07-19 |
DE19743518A1 (en) | 1999-04-15 |
CA2305171A1 (en) | 1999-04-08 |
EP1019430B1 (en) | 2007-07-11 |
KR20010024366A (en) | 2001-03-26 |
DK1019430T3 (en) | 2007-11-12 |
AU1028299A (en) | 1999-04-23 |
JP2001518284A (en) | 2001-10-16 |
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