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WO1993025709A1 - Preparation d'acides nucleiques - Google Patents

Preparation d'acides nucleiques Download PDF

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Publication number
WO1993025709A1
WO1993025709A1 PCT/GB1993/001223 GB9301223W WO9325709A1 WO 1993025709 A1 WO1993025709 A1 WO 1993025709A1 GB 9301223 W GB9301223 W GB 9301223W WO 9325709 A1 WO9325709 A1 WO 9325709A1
Authority
WO
WIPO (PCT)
Prior art keywords
sequence
complementary
probe
nucleic acid
oligonucleotides
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/GB1993/001223
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English (en)
Inventor
Trevor Leonard Hawkins
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Medical Research Council
Original Assignee
Medical Research Council
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Medical Research Council filed Critical Medical Research Council
Priority to AU43440/93A priority Critical patent/AU4344093A/en
Publication of WO1993025709A1 publication Critical patent/WO1993025709A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • 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
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L7/00—Heating or cooling apparatus; Heat insulating devices
    • B01L7/52—Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples
    • 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
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00277—Apparatus
    • B01J2219/00497—Features relating to the solid phase supports
    • B01J2219/005—Beads
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00718—Type of compounds synthesised
    • B01J2219/0072—Organic compounds
    • B01J2219/00722—Nucleotides
    • C—CHEMISTRY; METALLURGY
    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B40/00—Libraries per se, e.g. arrays, mixtures
    • C40B40/04—Libraries containing only organic compounds
    • C40B40/06—Libraries containing nucleotides or polynucleotides, or derivatives thereof
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/10—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices
    • G01N35/1081—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices characterised by the means for relatively moving the transfer device and the containers in an horizontal plane
    • G01N35/109—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices characterised by the means for relatively moving the transfer device and the containers in an horizontal plane with two horizontal degrees of freedom

Definitions

  • This invention relates to the preparation of nucleic acids and concerns magnetic particles and their use in the preparation of nucleic acids.
  • M13 phage is used to obtain single stranded DNA for sequencing templates.
  • the traditional methods for M13 DNA purification such as polyethylene glycol (PEG)/phenol procedures (Bankier et al . , [1988], in Wu, R. [ed.] , Methods Enz. 155, 52-93) allow microgram quantities of template to be produced from illilitre volumes.
  • thermally cycled sequencing procedures utilising Taq polymerase (Craxton, [1991], Methods: A Companion to Methods in Enzymology 3_, 20-26) only require 200-500ng of - template DNA per reaction. Therefore traditional methods produce an excess of template for most purposes, which wastes both time and money.
  • the invention provides a composition comprising magnetic particles, each bearing a plurality of oligonucleotides, each oligonucleotide comprising a sequence, acting as a probe, which is complementary to a sequence under investigation, and further comprising a sequence which is non-complementary to the sequence under investigation.
  • the invention provides a method of producing the composition defined above, comprising attaching magnetic particles to an oligonucleotide comprising a complementary probe sequence and further comprising a non-complementary sequence.
  • the magnetic particles take the form of beads. Typically these are about 1-5um in diameter. Suitable magnetic beads are commercially available. For instance, they may be obtained from Dynal or, more preferably, from Promega. Those from Promega are found particularly suitable because they have a pitted surface and therefore a higher surface area, which allows greater ' amounts of oligonucleotides to be carried by the bead.
  • 'Oligonucleotides' as used herein includes both synthetic and native DNA and RNA sequences of any length.
  • the oligonucleotide is preferably single stranded DNA and, conveniently, the complementary probe sequence is substantially 25-45 bases long.
  • the non-complementary sequence is generally 5-15 bases in length, typically about 9 bases long.
  • the non-complementary sequence of the oligonucleotide acts as a 'linker' joining the complementary probe sequence to the magnetic bead.
  • the probe sequence may be attached by either end to the linker oligonucleotide.
  • the oligonucleotide may be attached to the magnetic bead by conventional techniques.
  • the linker oligonucleotide may conveniently be biotinylated at the end region distal to the end joined to the complementary probe sequence, allowing for ready attachment to streptavidin-coated magnetic beads.
  • Other attachment techniques are known to those skilled in the art.
  • the probe sequence can, of course, be selected so as to be complementary to any nucleic acid sequence of interest such that, for example, the sequence of interest may be separated from a complex sample by being bound by the magnetic bead/probe and then separated from the rest of the sample by magnetic attraction.
  • the invention provides a method of separating a nucleic acid sequence from a sample comprising: contacting a complex sample possibly including the sequence of interest with the composition of the present invention; allowing the sequence of interest, if present, to associate with the composition by means of hybridisation with the complementary probe sequence; and separating the composition and any bound nucleic acid sequences from the rest of the sample by magnetic attraction.
  • the complementary probe sequence comprises an oligonucleotide complementary to the sequence of M13.
  • the invention therefore provides a method of preparing template DNA for sequencing.
  • the probe sequence is complementary to a region upstream from the M13 -21 universal primer site.
  • the arrangement described above can confer a number of advantages over prior art methods of purifying nucleic acids.
  • the use of a non-complementary linker sequence reduces the likelihood of steric hindrance between the comparatively bulky magnetic bead and the "target" DNA hybridised to the probe. This allows beads prepared in accordance with the invention to bind far more 'target' DNA than possible hitherto.
  • probe sequences may become separated from magnetic beads of the prior art by shearing and may act as unwanted "false" primers in subsequent sequencing reactions.
  • the non-complementary linker oligonucleotide of the invention ensures that, should the oligonucleotides become separated from the beads, they are far less likely to be able to prime chain-extension reactions. It is a preferred feature therefore that the linker sequence should be at the 3' end of the complementary probe sequence, as this further reduces the risk of non-specific priming.
  • the invention provides a method of preparing a DNA sequence of interest from a complex sample comprising: causing lysis of DNA-containing entities by the action of heat in the presence of a suitable detergent; adding magnetic particles attached to a probe complementary to at least part of the sequence of interest together with hybridisation buffer containing a polymer to cause an increase in the effective nucleic acid concentration; allowing the sequence of interest to hybridise to the complementary probe and then separating the particle/probe complex and any sequences hybridised thereto from the rest of the sample by magnetic attraction.
  • any magnetic particles to which are attached an appropriate probe are suitable for performing the method defined above, including those already known in the art. However, it is preferred that the particles used are those according to the present invention.
  • a suitable detergent is SDS.
  • the DNA-containing entities are typically phages.
  • a suitable polymer is PEG.
  • the use of a polymer to increase the effective concentration of nucleic acid allows smaller volumes of reagents to be used than previously, which in turn enables the method to be performed in a microtitre plate or tray. Similarly, the method is particularly simple and efficient as it employs a target sequence-specific purification step.
  • the invention provides a method of preparing DNA by means of a sequence-specific purification step, capable of being performed in a microtitre plate.
  • the ability to perform the method in a microtitre plate makes the method of the invention particularly amenable to automation.
  • Suitable automated apparatus for performing the method of the invention is described in GB9212164.9 (co-pending Application No PCT/GB93/ ).
  • Figure 1 shows the base sequence of a suitable probe/linker oligonucleotide
  • Figure 2 shows a photograph of gel electrophoresis performed on DNA prepared by the method of the invention.
  • Figure 3 shows a portion of sequencing trace
  • the invention involves an oligonucleotide probe which has been synthesised with a biotin group at the 3' end.
  • the probe is designed to be complementary to a region upstream from the M13 -21 Universal priming site.
  • single plaques may be grown up in culture, the cells harvested and the supernatant collected and lysed to yield free single strands.
  • the bead/probe complex is then added, and the probe allowed to anneal to the target DNA. Once bound, the bead/probe/template complex can be separated from the rest of the sample using magnetic attraction and then washed.
  • the template may then be freed from the bead/probe complex by heating.
  • the procedure is simple and fast. All the post-growth steps can be carried out in microtitre plates with no centrifugation or ethanol precipitations required. In this example, 250 random M13 sub-clones were prepared and se ⁇ uenced using the method outlined below.
  • the amount of template recovered depends directly on the design of the probe.
  • the probe must be specific to the target but must not act as a secondary sequencing primer if free probe is left in solution. Consequently, the probe is 41bp in length and binds to a region upstream from the M13 -21 Universal primer site.
  • the probe has a run of several 'A's at the 3' end together with the biotin group. This acts as a linker arm to prevent steric hindrance between the large streptavidin-coated beads and the binding of the probe to the target. Also the high degree of non-complementarity at the 3 * end would prevent the free probe from acting as a sequencing primer should it shear from the beads.
  • the design of the probe is shown in Figure 1, ('B' represents Biotin).
  • the probe THM13.3 was synthesised on an ABI 380B DNA synthesiser on a 1uMole scale. Biotin phosphoramidite was obtained frm Amersham U.K. The sequence of the probe was: 5' TAT CGG CCT CAG GAA GAT CGC ACT CCA GCC AGC AAA AAA Biotin A 3' (Seq ID No. 1). Following cleavage from the column, the oligonucleotide was deprotected in ammonia at 55°C overnight. A NAP-10 column was used to purify the crude oligonucleotide. Probe/streptavidin bead linkage
  • Promega nucleotide quality beads were used in this example. 1.2ml of Promega beads were used per 12 samples. The beads were washed in 0.1M NaCl three times using a neodymium-iron-boron permanent magnet to separate the beads from the washing solution. 200ul 0.1M NaCl and 1On ol THM13.3 oligonucleotide were then added to 1.2ml( 1.2mg) dry beads and incubated at room temperature for 10 minutes. The beads were then washed 10 times in 0.1M NaCl to remove unbound oligonucleotide. Bead/probe complex was finally taken up in 1.2ml water. Beads may be bound to probe in bulk and stored in storage buffer at 4°C.
  • Random M13 sub-clones with 1-2kb inserts were grown up in 2ml 2xTy medium (16g bacto tryptone, 10g yeast extract, 5g NaCl in 11 water) for 5 hours at 37°C. Cells were spun down at 14,000g for 5 minutes and the supernatant transferred to microtitre plates. 400ul supernatant was used from each sample, using two wells, each containing 200ul supernatant (Falcon 3911 MicroTest Flexible Assay Plate. )
  • the microtitre plate was removed from the cycler and placed on a Dynal MPC-96 magnetic separator. After 30 seconds the supernatant was aspirated, 100ul wash buffer (0.1X SSC) were added to each well and the beads were moved through the wash buffer by repositioning the plate over the magnetic separator three times at intervals of five seconds. This step was repeated a total of three times. Finally, the beads were eluted in 10ul water; using the magnet, beads may be dispersed into this small volume.
  • 100ul wash buffer 0.1X SSC
  • the templates were released from the probe by heating the plate to 80 C for 3 minutes. After heating, the beads were concentrated using the magnet and the supernatant was removed to a fresh microtitre plate. Due to evaporation the final total volume was approximately 8ul from each well, or approximately 16ul per DNA template sample.
  • FIG. 2 The efficacy of the technique is illustrated by Figure 2.
  • Four random M13 subclones were grown as described. The cells were harvested and the supernatant (1200ul) was split three ways to provide three identical samples. A, B and C (400ul each). For each sub clone. Sample A was prepared using the method outlined previously. Sample B was prepared as outlined but leaving out the hybridisation buffer (20% PEG/2.5M NaCl). Sample C was prepared as outlined but using magnetic beads without a probe attached. The samples were then subjected to agarose gel electrophoresis. This showed that when the PEG is removed there is a dramatic reduction in yield, as expected. When beads are used without probe attached the resulting yield is zero. The difference between this control and the method of Alderton et al., (1992), is that in sample C the phage are lysed before addition of PEG.
  • beads may be re-used as outlined below.
  • Beads were collected and pooled in a 1.5ml eppendorf tube. The supernatant was removed and the beads were resuspended in 1ml reuse buffer (0.15M NaOH, 0.001% Tween 20) for 1 minute. The supernatant was removed and the procedure repeated. Finally the beads were washed once in storage buffer and taken up in half the original volume of storage buffer (PBS pH 7.5, 0.1% BSA). Beads were then stored at 4°C.
  • a preferred feature of the method of preparing DNA is to bring about magnetic attraction for the magnetic beads by the use of a "dot magnet".
  • a typical microtitre assay plate generally 130 mm by 85 mm
  • the dot magnet comprises an array of 96 such rare earth magnets which are positioned in a matrix 8 x 12 and are separated by about 9mm from each nearest neighbour (ie. substantially the same spacing as that between the wells of a microtitre plate) thus when the sides of the dot magnet are aligned with the sides of a typical microtitre plate, each rare earth magnet is positioned beneath a well in the microtitre plate.
  • MOLECULE TYPE DNA (genomic)
  • HYPOTHETICAL NO
  • ANTI-SENSE NO

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Abstract

L'invention se rapporte à une composition comprenant des particules magnétiques, dont chacune porte une multiplicité d'oligonucléotides, chaque nucléotide comprenant une séquence agissant comme une sonde, qui est complémentaire d'une séquence étudiée, et comprenant en outre une séquence qui n'est pas complémentaire de la séquence étudiée. Un procédé de purification d'acides nucléiques à l'aide d'une composition de l'invention est également décrit, ainsi qu'un procédé de préparation d'un acide nucléique.
PCT/GB1993/001223 1992-06-09 1993-06-09 Preparation d'acides nucleiques Ceased WO1993025709A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU43440/93A AU4344093A (en) 1992-06-09 1993-06-09 Preparation of nucleic acids

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GB929212164A GB9212164D0 (en) 1992-06-09 1992-06-09 Preparation of nucleic acids
GB9212164.9 1992-06-09

Publications (1)

Publication Number Publication Date
WO1993025709A1 true WO1993025709A1 (fr) 1993-12-23

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Application Number Title Priority Date Filing Date
PCT/GB1993/001222 Ceased WO1993025912A2 (fr) 1992-06-09 1993-06-09 Preparation automatique d'acides nucleiques
PCT/GB1993/001223 Ceased WO1993025709A1 (fr) 1992-06-09 1993-06-09 Preparation d'acides nucleiques

Family Applications Before (1)

Application Number Title Priority Date Filing Date
PCT/GB1993/001222 Ceased WO1993025912A2 (fr) 1992-06-09 1993-06-09 Preparation automatique d'acides nucleiques

Country Status (3)

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AU (2) AU4343993A (fr)
GB (1) GB9212164D0 (fr)
WO (2) WO1993025912A2 (fr)

Cited By (27)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1996009379A1 (fr) * 1994-09-20 1996-03-28 Whitehead Institute For Biomedical Research Purification et isolation d'adn au moyen d'une phase solide
US5683875A (en) * 1995-05-04 1997-11-04 Hewlett-Packard Company Method for detecting a target nucleic acid analyte in a sample
DE19801730A1 (de) * 1998-01-19 1999-07-22 Fraunhofer Ges Forschung Verfahren zur kombinierten Analyse von Zellinhalten, insbesondere den in biologischen Zellen enthaltenen Nukleinsäuren sowie Vorrichtung zur Durchführung des Verfahrens
DE19943374A1 (de) * 1999-09-10 2001-03-29 Max Planck Gesellschaft Verfahren zum Anbinden von Nukleinsäuren an eine Festphase
WO2001051665A3 (fr) * 2000-01-13 2003-01-30 Nanosphere Inc Nanoparticules auxquelles sont attaches des oligonucleotides et utilisations de ces dernieres
WO2001073123A3 (fr) * 2000-03-28 2003-02-06 Nanosphere Inc Nanoparticules auxquelles sont attaches des oligonucleotides et leur utilisation
US6582921B2 (en) 1996-07-29 2003-06-24 Nanosphere, Inc. Nanoparticles having oligonucleotides attached thereto and uses thereof
US6602669B2 (en) 2000-07-11 2003-08-05 Northwestern University Method of detection by enhancement of silver staining
WO2002046472A3 (fr) * 2000-12-08 2003-09-04 Nanosphere Inc Nanoparticules auxquelles sont attaches des oligonucleotides et utilisations de ces dernieres
WO2002018643A3 (fr) * 2000-08-11 2003-09-04 Nanosphere Inc Nanoparticules comportant des oligonucleotides fixes dessus et utilisations associees
US6645721B2 (en) 1996-07-29 2003-11-11 Nanosphere, Inc. Nanoparticles having oligonucleotides attached thereto and uses therefor
US6750016B2 (en) 1996-07-29 2004-06-15 Nanosphere, Inc. Nanoparticles having oligonucleotides attached thereto and uses therefor
US6773884B2 (en) 1996-07-29 2004-08-10 Nanosphere, Inc. Nanoparticles having oligonucleotides attached thereto and uses therefor
US6812334B1 (en) 1996-07-29 2004-11-02 Nanosphere, Inc. Nanoparticles having oligonucleotides attached thereto and uses therefor
US6974669B2 (en) 2000-03-28 2005-12-13 Nanosphere, Inc. Bio-barcodes based on oligonucleotide-modified nanoparticles
US6977178B2 (en) 2000-09-06 2005-12-20 Transnetyx, Inc. System and method for transgenic and targeted mutagenesis screening
US6984491B2 (en) 1996-07-29 2006-01-10 Nanosphere, Inc. Nanoparticles having oligonucleotides attached thereto and uses therefor
WO2006059386A1 (fr) * 2004-12-02 2006-06-08 National Institute Of Agrobiological Sciences Procédé consistant à isoler un adn double brin contenant des séquences répétées en tandem (ssr)
US7115688B1 (en) 1998-11-30 2006-10-03 Nanosphere, Inc. Nanoparticles with polymer shells
US7135055B2 (en) 2001-05-25 2006-11-14 Nanosphere, Inc. Non-alloying core shell nanoparticles
US7147687B2 (en) 2001-05-25 2006-12-12 Nanosphere, Inc. Non-alloying core shell nanoparticles
US7169556B2 (en) 1996-07-29 2007-01-30 Nanosphere, Inc. Nanoparticles having oligonucleotides attached thereto and uses therefor
US7186814B2 (en) 2001-11-09 2007-03-06 Nanosphere, Inc. Bioconjugate-nanoparticle probes
US7253277B2 (en) 2002-07-02 2007-08-07 Nanosphere, Inc. Nanoparticle polyanion conjugates and methods of use thereof in detecting analytes
US7494817B2 (en) 2000-09-06 2009-02-24 Transnet Yx, Inc. Methods for genotype screening using magnetic particles
US7527929B2 (en) 2004-07-30 2009-05-05 Agencourt Bioscience Corporation Methods of isolating nucleic acids using multifunctional group-coated solid phase carriers
US8691969B2 (en) 1994-12-12 2014-04-08 Life Technologies As Isolation of nucleic acid

Families Citing this family (30)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE4409436A1 (de) * 1994-03-19 1995-09-21 Boehringer Mannheim Gmbh Verfahren zur Bearbeitung von Nukleinsäuren
DE4421058A1 (de) 1994-06-16 1995-12-21 Boehringer Mannheim Gmbh Verfahren zur magnetischen Abtrennung von Flüssigkeitskomponenten
CN1170460A (zh) * 1994-10-07 1998-01-14 弗林德斯科技有限公司 用于生物多聚体的储存、提纯或反应以及处理的装置和方法
DE19512368A1 (de) * 1995-04-01 1996-10-02 Boehringer Mannheim Gmbh System zur Freisetzung und Isolierung von Nukleinsäuren
US6919175B1 (en) 1995-04-01 2005-07-19 Roche Diagnostics Gmbh System for releasing and isolating nucleic acids
US5779985A (en) * 1995-12-18 1998-07-14 Solid Phase Sciences Corporation Reaction plenum
GB9525794D0 (en) * 1995-12-18 1996-02-21 Hale Alan Biotechnological process
US6277332B1 (en) 1995-12-18 2001-08-21 Solid Phase Sciences Corporation Reaction plenum with magnetic separation and/or ultrasonic agitation
US6008010A (en) 1996-11-01 1999-12-28 University Of Pittsburgh Method and apparatus for holding cells
FR2760844B1 (fr) * 1997-03-17 1999-05-21 Genethon Ii Dispositif et procede d'analyse automatique d'echantillons sur gels
US5912129A (en) * 1998-03-05 1999-06-15 Vinayagamoorthy; Thuraiayah Multi-zone polymerase/ligase chain reaction
AU2003271382B2 (en) * 1998-05-01 2006-07-20 Gen-Probe Incorporated Automated Diagnostic Analyzer and Method
AU2006230729B2 (en) * 1998-05-01 2008-04-03 Gen-Probe Incorporated Transport mechanism
US8337753B2 (en) 1998-05-01 2012-12-25 Gen-Probe Incorporated Temperature-controlled incubator having a receptacle mixing mechanism
DE10012007T1 (de) * 1998-05-01 2012-01-26 Gen-Probe Inc. Automatisches Diagnoseanalysegerät und Verfahren
DE19854003A1 (de) * 1998-11-18 2000-05-25 Jenoptik Jena Gmbh Simultanes Magnetpartikelhandling in zweidimensionaler Anordnung
CA2378573A1 (fr) * 1999-07-05 2001-01-11 Max-Planck-Gesellschaft Zur Forderung Der Wissenschaften E.V. Procede de selection a haut rendement de partenaires de liaison
DE19954840A1 (de) * 1999-11-09 2001-05-17 Biopsytec Gmbh Vorrichtung zur Aufbereitung biologischer Proben für die DNA-Analyse
JP2002022752A (ja) * 2000-07-13 2002-01-23 Suzuki Motor Corp 検体試験装置
DE10065148B4 (de) * 2000-12-22 2004-01-22 Institut für Physikalische Hochtechnologie e.V. Magnetische Greif-Halte-Anordnung
CA2842404C (fr) 2005-03-10 2016-05-10 Gen-Probe Incorporated Systemes et procedes permettant d'effectuer des dosages afin de detecter ou de quantifier des analytes dans des echantillons
EP2752247A3 (fr) 2010-07-23 2016-08-31 Beckman Coulter, Inc. Système ou procédé comprenant des unités analytiques
US9046507B2 (en) 2010-07-29 2015-06-02 Gen-Probe Incorporated Method, system and apparatus for incorporating capacitive proximity sensing in an automated fluid transfer procedure
CN103403533B (zh) 2011-02-24 2017-02-15 简.探针公司 用于分辨光信号检测器中不同调制频率的光信号的系统和方法
WO2013070754A1 (fr) 2011-11-07 2013-05-16 Beckman Coulter, Inc. Bras robotique
WO2013070748A1 (fr) 2011-11-07 2013-05-16 Beckman Coulter, Inc. Amortissement magnétique pour système de transport d'échantillon
US9482684B2 (en) 2011-11-07 2016-11-01 Beckman Coulter, Inc. Centrifuge system and workflow
WO2013070740A1 (fr) 2011-11-07 2013-05-16 Beckman Coulter, Inc. Système d'aliquote et flux de travail
BR112014010955A2 (pt) 2011-11-07 2017-06-06 Beckman Coulter Inc sistema e método para processar amostras
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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1986005815A1 (fr) * 1985-03-25 1986-10-09 Genetics International Inc. Sequences d'acide nucleique liees a des materiaux sensibles a des champs magnetiques, et methodes d'analyse et appareils utilisant lesdites sequences liees
WO1990006045A2 (fr) * 1988-11-21 1990-06-14 Dynal As Sondes d'acide nucleique

Family Cites Families (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AU596987B2 (en) * 1985-08-30 1990-05-24 Tosoh Corporation Automated immunoassay analyser
FR2633310A1 (fr) * 1988-06-24 1989-12-29 Pasteur Institut Appareil d'execution automatique repetee d'un cycle thermique en plusieurs etapes successives, notamment pour l'amplification enzymatique de sequences d'acides nucleiques
GB8816982D0 (en) * 1988-07-16 1988-08-17 Probus Biomedical Ltd Bio-fluid assay apparatus
CA2031912A1 (fr) * 1989-12-22 1991-06-23 Robert Fred Pfost Couvercle chauffe
EP0478753B1 (fr) * 1990-04-06 1997-07-02 The Perkin-Elmer Corporation Laboratoire de biologie moleculaire automatise
WO1992005443A1 (fr) * 1990-09-15 1992-04-02 Medical Research Council Separation de reactif
EP0479448A3 (en) * 1990-10-02 1992-12-23 Beckman Instruments, Inc. Magnetic separation device

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1986005815A1 (fr) * 1985-03-25 1986-10-09 Genetics International Inc. Sequences d'acide nucleique liees a des materiaux sensibles a des champs magnetiques, et methodes d'analyse et appareils utilisant lesdites sequences liees
WO1990006045A2 (fr) * 1988-11-21 1990-06-14 Dynal As Sondes d'acide nucleique

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
ANALYTICAL BIOCHEMISTRY vol. 201, February 1992, NEW YORK US pages 166 - 169 ALDERTON ET AL. 'Magnetic bead purification of M13 DNA sequencing templates' cited in the application *

Cited By (59)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1996009379A1 (fr) * 1994-09-20 1996-03-28 Whitehead Institute For Biomedical Research Purification et isolation d'adn au moyen d'une phase solide
US5705628A (en) * 1994-09-20 1998-01-06 Whitehead Institute For Biomedical Research DNA purification and isolation using magnetic particles
US5898071A (en) * 1994-09-20 1999-04-27 Whitehead Institute For Biomedical Research DNA purification and isolation using magnetic particles
US8691969B2 (en) 1994-12-12 2014-04-08 Life Technologies As Isolation of nucleic acid
US5683875A (en) * 1995-05-04 1997-11-04 Hewlett-Packard Company Method for detecting a target nucleic acid analyte in a sample
US6818753B2 (en) 1996-07-29 2004-11-16 Nanosphere, Inc. Nanoparticles having oligonucleotides attached thereto and uses therefor
US6750016B2 (en) 1996-07-29 2004-06-15 Nanosphere, Inc. Nanoparticles having oligonucleotides attached thereto and uses therefor
US6986989B2 (en) 1996-07-29 2006-01-17 Nanosphere, Inc. Nanoparticles having oligonucleotides attached thereto and uses therefor
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US6902895B2 (en) 1996-07-29 2005-06-07 Nanosphere, Inc. Nanoparticles having oligonucleotides attached thereto and uses therefor
US6610491B2 (en) 1996-07-29 2003-08-26 Nanosphere, Inc. Nanoparticles having oligonucleotides attached thereto and uses therefor
US7259252B2 (en) 1996-07-29 2007-08-21 Nanosphere, Inc. Nanoparticles having oligonucleotides attached thereto and uses therefor
US7250499B2 (en) 1996-07-29 2007-07-31 Nanosphere Inc. Nanoparticles having oligonucleotides attached thereto and uses therefor
US6645721B2 (en) 1996-07-29 2003-11-11 Nanosphere, Inc. Nanoparticles having oligonucleotides attached thereto and uses therefor
US6903207B2 (en) 1996-07-29 2005-06-07 Nanosphere, Inc. Nanoparticles having oligonucleotides attached thereto and uses therefor
US6677122B2 (en) 1996-07-29 2004-01-13 Nanosphere, Inc. Nanoparticles having oligonucleotides attached thereto and uses therefor
US6682895B2 (en) 1996-07-29 2004-01-27 Nanosphere, Inc. Nanoparticles having oligonucleotides attached thereto and uses therefor
US6709825B2 (en) 1996-07-29 2004-03-23 Nanosphere, Inc. Nanoparticles having oligonucleotides attached thereto and uses therefor
US6720147B2 (en) 1996-07-29 2004-04-13 Nanosphere, Inc. Nanoparticles having oligonucleotides attached thereto and uses therefor
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US7098320B1 (en) 1996-07-29 2006-08-29 Nanosphere, Inc. Nanoparticles having oligonucleotides attached thereto and uses therefor
US6759199B2 (en) 1996-07-29 2004-07-06 Nanosphere, Inc. Nanoparticles having oligonucleotides attached thereto and uses therefor
US6767702B2 (en) 1996-07-29 2004-07-27 Nanosphere, Inc. Nanoparticles having oligonucleotides attached thereto and uses therefor
US6773884B2 (en) 1996-07-29 2004-08-10 Nanosphere, Inc. Nanoparticles having oligonucleotides attached thereto and uses therefor
US6777186B2 (en) 1996-07-29 2004-08-17 Nanosphere, Inc. Nanoparticles having oligonucleotides attached thereto and uses therefor
US6812334B1 (en) 1996-07-29 2004-11-02 Nanosphere, Inc. Nanoparticles having oligonucleotides attached thereto and uses therefor
US6984491B2 (en) 1996-07-29 2006-01-10 Nanosphere, Inc. Nanoparticles having oligonucleotides attached thereto and uses therefor
US6828432B2 (en) 1996-07-29 2004-12-07 Nanosphere, Inc. Nanoparticles having oligonucleotides attached thereto and uses therefor
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US7208587B2 (en) 1996-07-29 2007-04-24 Nanosphere, Inc. Nanoparticles having oligonucleotides attached thereto and uses therefor
US6673548B2 (en) 1996-07-29 2004-01-06 Nanosphere, Inc. Nanoparticles having oligonucleotides attached thereto and uses therefor
US6962786B2 (en) 1996-07-29 2005-11-08 Nanosphere, Inc. Nanoparticles having oligonucleotides attached thereto and uses therefor
US6969761B2 (en) 1996-07-29 2005-11-29 Nanosphere, Inc. Nanoparticles having oligonucleotides attached thereto and uses therefor
US7169556B2 (en) 1996-07-29 2007-01-30 Nanosphere, Inc. Nanoparticles having oligonucleotides attached thereto and uses therefor
DE19801730A1 (de) * 1998-01-19 1999-07-22 Fraunhofer Ges Forschung Verfahren zur kombinierten Analyse von Zellinhalten, insbesondere den in biologischen Zellen enthaltenen Nukleinsäuren sowie Vorrichtung zur Durchführung des Verfahrens
US7115688B1 (en) 1998-11-30 2006-10-03 Nanosphere, Inc. Nanoparticles with polymer shells
DE19943374A1 (de) * 1999-09-10 2001-03-29 Max Planck Gesellschaft Verfahren zum Anbinden von Nukleinsäuren an eine Festphase
WO2001051665A3 (fr) * 2000-01-13 2003-01-30 Nanosphere Inc Nanoparticules auxquelles sont attaches des oligonucleotides et utilisations de ces dernieres
WO2001073123A3 (fr) * 2000-03-28 2003-02-06 Nanosphere Inc Nanoparticules auxquelles sont attaches des oligonucleotides et leur utilisation
US6974669B2 (en) 2000-03-28 2005-12-13 Nanosphere, Inc. Bio-barcodes based on oligonucleotide-modified nanoparticles
US7323309B2 (en) 2000-03-28 2008-01-29 Northwestern University Bio-barcodes based on oligonucleotide-modified particles
US6602669B2 (en) 2000-07-11 2003-08-05 Northwestern University Method of detection by enhancement of silver staining
WO2002018643A3 (fr) * 2000-08-11 2003-09-04 Nanosphere Inc Nanoparticules comportant des oligonucleotides fixes dessus et utilisations associees
US7011943B2 (en) 2000-09-06 2006-03-14 Transnetyx, Inc. Method for detecting a designated genetic sequence in murine genomic DNA
US6977178B2 (en) 2000-09-06 2005-12-20 Transnetyx, Inc. System and method for transgenic and targeted mutagenesis screening
US7494817B2 (en) 2000-09-06 2009-02-24 Transnet Yx, Inc. Methods for genotype screening using magnetic particles
US7282361B2 (en) 2000-09-06 2007-10-16 Transnetyx, Inc. Systems and methods for transgenic and targeted mutagenesis screening
WO2002046472A3 (fr) * 2000-12-08 2003-09-04 Nanosphere Inc Nanoparticules auxquelles sont attaches des oligonucleotides et utilisations de ces dernieres
US7147687B2 (en) 2001-05-25 2006-12-12 Nanosphere, Inc. Non-alloying core shell nanoparticles
US7238472B2 (en) 2001-05-25 2007-07-03 Nanosphere, Inc. Non-alloying core shell nanoparticles
US7135055B2 (en) 2001-05-25 2006-11-14 Nanosphere, Inc. Non-alloying core shell nanoparticles
US7186814B2 (en) 2001-11-09 2007-03-06 Nanosphere, Inc. Bioconjugate-nanoparticle probes
US7253277B2 (en) 2002-07-02 2007-08-07 Nanosphere, Inc. Nanoparticle polyanion conjugates and methods of use thereof in detecting analytes
US7527929B2 (en) 2004-07-30 2009-05-05 Agencourt Bioscience Corporation Methods of isolating nucleic acids using multifunctional group-coated solid phase carriers
JPWO2006059386A1 (ja) * 2004-12-02 2008-06-05 独立行政法人農業生物資源研究所 単純反復配列を含む２本鎖ｄｎａの単離方法
WO2006059386A1 (fr) * 2004-12-02 2006-06-08 National Institute Of Agrobiological Sciences Procédé consistant à isoler un adn double brin contenant des séquences répétées en tandem (ssr)

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WO1993025912A3 (fr) 1994-03-17
AU4344093A (en) 1994-01-04
AU4343993A (en) 1994-01-04
WO1993025912A2 (fr) 1993-12-23
GB9212164D0 (en) 1992-07-22

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