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WO1996000228A1 - Procede d'isolation rapide d'acide nucleique - Google Patents

Procede d'isolation rapide d'acide nucleique Download PDF

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Publication number
WO1996000228A1
WO1996000228A1 PCT/US1995/007940 US9507940W WO9600228A1 WO 1996000228 A1 WO1996000228 A1 WO 1996000228A1 US 9507940 W US9507940 W US 9507940W WO 9600228 A1 WO9600228 A1 WO 9600228A1
Authority
WO
WIPO (PCT)
Prior art keywords
nucleic acid
protein
lysing buffer
rna
amplification
Prior art date
Application number
PCT/US1995/007940
Other languages
English (en)
Inventor
Janice T. Brown
Original Assignee
Dade International Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Dade International Inc. filed Critical Dade International Inc.
Priority to AU29079/95A priority Critical patent/AU2907995A/en
Publication of WO1996000228A1 publication Critical patent/WO1996000228A1/fr

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/08—Reducing the nucleic acid content

Definitions

  • This invention relates to isolation of nucleic acids from biological samples. More
  • this invention relates to methods and articles of manufacture useful for
  • RNA or DNA isolating RNA or DNA from biological samples in a simple, rapid manner and to the use
  • Isolated nucleic acids are useful for many biotechnology-related purposes such as gene cloning, analysis of translation products in vitro and detection of particular sequences for diagnostic assays. Isolated nucleic acids often serve as
  • PCR polymerase chain reaction
  • RNA ribonucleic acid
  • RNA that can be translated in an in vitro translation system and that is suitable for
  • RNA isolation is the presence of ribonucleases in
  • RNA generally results in unwanted degradation of RNA species present in the sample. Ribonucleases are noted for their stability under conditions that denature most other
  • RNA is preventing ribonuclease activity
  • DNA isolation such as known methods for rapid isolation of plasmid DNA from
  • nucleic acid isolation methods suffer from one or more of the known nucleic acid isolation methods.
  • nucleic acids for example, for use in a clinical assay.
  • a clinical work-up of a biological sample in which nucleic acid will be analyzed generally requires that nucleic
  • RNA comprising RNA, DNA and protein, in which the isolated RNA is substantially intact.
  • the method comprises the steps of a) incubating the sample in a lysing buffer comprising
  • nucleic acid from the nucleic acid solution using a nucleic acid-precipitating agent
  • the lysing buffer is substantially free of
  • guanidine compounds such as guanidine thiocyanate.
  • the lysing buffer also may include a proteinase or a ribonuclease inhibitor.
  • ribonuclease inhibitor may comprise vanadyl ribonucleoside complex.
  • the incubating step may occur for a time from less than about 1 minute to about
  • the method may further comprise
  • Nucleic acid recovered according to the invention may be suitable for use as a
  • recovered RNA may be effective for
  • cDNA synthesis and amplification by 3SR and recovered DNA may be effective for
  • An article of manufacture comprising packaging material and a
  • the lysing buffer comprises an ionic detergent, and is substantially free of guanidine compounds.
  • the article further comprises a label or package insert accompanying the packaging material, which
  • lysing buffer indicates that the lysing buffer is suitable for isolating total nucleic acid according to a
  • An article of manufacture may further comprise a salt
  • composition in which case the label or package insert further indicates that the salt
  • composition can be used to isolate total nucleic acid according to a method of the
  • Figure 1 is a photograph of an ethidium bromide-stained agarose gel showing the yield of nucleic acid isolated according to the invention.
  • Figure 2 is a photograph of an ethidium-bromide stained agarose gel. Lanes 1
  • Lanes 2-5 show l/20th of the nucleic acid
  • Figure 3 is an autoradiogram of a slot blot, showing 3SR reaction products prepared from template nucleic acid isolated in Figure 1. Slots A1-A3 show the products
  • Dl shows the product prepared from nucleic acid in lane 12 of Figure 1.
  • Slot El shows the product prepared from nucleic acid in lane 12 of Figure 1.
  • Figure 4 is a photograph of an ethidium bromide-stained agarose gel showing the
  • Figure 5 is a photograph of an ethidium bromide-stained agarose gel showing the
  • nucleic acid yield using different combinations of salt solutions and nucleic acid
  • Figure 6 is an autoradiogram of a Northern blot of nucleic acid samples isolated
  • Lanes 4-6 1, 5, and 10 ⁇ l of nucleic acid isolated without proteinase K in
  • Figure 7 is a photograph of an ethidium bromide-stained agarose gel, showing
  • Lane 1 0X174
  • Lanes 9-10 2 and 7 ⁇ l respectively of nucleic acid from K562 cells converted
  • Lane 11 7 ⁇ l of nucleic acid from K562 cells amplified by PCR using primers BB160 and BB165
  • Lane 12 7 ⁇ l of nucleic acid from K562 cells isolated by
  • RNAzol B converted to first strand cDNA and amplified by PCR using primers BB164 and BB165; Lane 13: 7 ytl of nucleic acid from K562 cells isolated by RNAzol B,
  • Lane 14 0X174 digested with Haelll.
  • Figure 8 is a photograph of an ethidium-bromide stained agarose gel. Lanes 1
  • Lanes 2-5 show different dilutions of the
  • Lane 2 1 :1000 dilution; lane 3, 1:750 dilution; lane 4, 1:500
  • Figure 9 is a photograph of an ethidium bromide-stained agarose gel, showing
  • nucleic acid from a biological sample, while utilizing few solution components
  • Methods of theinvention involve lysing a biological
  • RNA present in the isolated nucleic acid Surprisingly, RNA present in the isolated nucleic acid
  • isolated DNA or RNA can be used as a template in an amplification reaction.
  • isolated nucleic acid is useful as a template for 3SR or PCR.
  • RNA isolated by the method of the invention is suitable for cloning of
  • An advantage of the present invention is the rapidity with which nucleic acid
  • the acid may be isolated from a biological sample.
  • the method of the invention shortens
  • a biological sample may comprise intact cells, clumps of cells, portions of cells, isolated nuclei, or tissue.
  • a biological sample may be preserved, e.g., fixed, frozen
  • RNA and DNA substantially intact RNA and DNA.
  • a biological sample may be derived
  • a biological sample may be fixed and/or frozen to allow later analysis, e.g.,
  • a biological sample may be a fresh preparation, i.e, a specimen that is to
  • a biological sample may be from a source such as animal tissue or cells,
  • tissue pieces including without limitation a cell culture, intact tissue pieces (e.g., a biopsy), or a whole
  • a biological sample may comprise plant or fungal tissue or cells, which may be processed before performing a method of the invention, for example, by
  • the amount of tissue or the number of cells present in a biological sample may be adjusted as desired, e.g., to achieve a desired yield of nucleic acid. For example, if cultured mammalian cells are the source of a biological sample and
  • the isolated nucleic acid is to be a template for an amplification reaction, a suitable
  • amount may be from about 500 to about 1 X 10 7 cells.
  • a biological sample is incubated in a lysing buffer, which comprises an ionic
  • SDS sodium dodecyl sulfate
  • the concentration of ionic detergent in a lysing buffer is
  • lysing buffer may be included in a lysing buffer if desired.
  • lysing buffer may comprise an ionic detergent, a ribonuclease inhibitor, a
  • lysing buffer may comprise an ionic detergent and a pH buffering agent.
  • additional components such as a sulffiydryl reducing agent and a
  • ribonuclease inhibitor do not interfere with isolation of DNA.
  • Ribonuclease inhibitors are optionally included in a lysing buffer to reduce
  • a ribonuclease inhibitor is vanadyl-ribonucleoside complex
  • Vanadyl-ribonucleoside complex comprises a mixture of the complexes formed between an oxovanadium IV ion and each of the four ribonucleosides.
  • suitableribonuclease inhibitors include malacoid and bentonite.
  • a sulffiydryl reducing agent optionally is included in the lysing buffer.
  • Illustrative examples of such reducing agents are dithiothreitol and ⁇ -mercaptoethanol.
  • a pH buffering agent is optionally included to ensure that no unwanted changes occur in
  • buffering agents include Tris (hydroxymethyl)-aminomethane, sodium or potassium
  • Lysing buffer optionally contains a proteinase, which may assist in freeing nuclear
  • Lysing buffer preferably contains a proteinase if a
  • biological sample comprises cell clumps or intact tissue pieces. Inclusion of a proteinase
  • nucleic acid is particularly preferred when the resulting nucleic acid is to be used as a template in an
  • proteinases are proteinase K, PronaseTM
  • Such proteinases typically are present in a lysing buffer from
  • a lysing buffer according to the invention is substantially free of guanidine
  • RNA isolation have guanidine compounds as a component.
  • the lysing has guanidine compounds as a component.
  • guanidine compounds refers to guanidine and guanidinium compounds and salts thereof, including, but not limited to
  • cells generally require from less than about 1 to about 120 minutes, preferably about 1 to
  • sample is sufficient to allow separation of protein and recovery of nucleic acids in
  • the amount of lysing buffer is adjusted according to factors such as
  • a solution comprising a major salt and a minor salt is then added to the incubated
  • a major salt may be, for example, potassium acetate, potassium chloride,
  • a minor salt may be a salt such as magnesium
  • magnesium ions are preferably used to form the salt composition since sodium salts are not as effective at precipitating SDS.
  • the amount of salt composition added to the incubated mixture is sufficient to provide
  • detergent-protein complexes to precipitate, e.g., in about 10X to about 100X
  • the concentration of the major salt is generally from
  • concentration of the minor salt is generally from about 25 niM to about 75 mM
  • complexes may be accelerated, if desired, by chilling the mixture on ice for a brief period
  • Precipitated detergent-protein is separated from the supernatant, which contains
  • detergent-protein may be any suitable means.
  • detergent-protein may be any suitable means.
  • detergent-protein may be any suitable means.
  • detergent-protein may be any suitable means.
  • detergent-protein may be any suitable means.
  • StratacleanTM resin (Stratagene, La Jolla, California, 92037), Pro-CipitateTM (Affinity Technology, Inc. New Brunswick, NY 08901) or other commercially available protein
  • detergent-protein may be separated by adding phenol and
  • the supernatant may
  • Phenol is a known protein-extracting agent that is
  • ribonuclease inhibitors such as vanadyl-ribonucleoside complex
  • 8-hydroxyquinoline may be performed, if desired.
  • nucleic acid-precipitating agent Suitable nucleic acid-precipitating agent
  • agents include 2 volumes of ethanol, 1/2 to 1 volume of isopropanol, or 1 volume of a
  • CTAB cetyltrimethylammoniurn bromide
  • the precipitated nucleic acid is recovered by means appropriate to the end use of
  • nucleic acids may be recovered by centrifuging the mixture in order to pellet the nucleic acid precipitate.
  • nucleic acids may be recovered by centrifuging the mixture in order to pellet the nucleic acid precipitate.
  • nucleic acids are examples of compounds that are useful in the art. They are: acetyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-methyl methyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl
  • the recovered nucleic acid is useful for a number of end uses.
  • the nucleic acid may be used to prepare genomic libraries or to clone
  • a particularly preferred use is as a template in an amplification protocol such as
  • the amount of HPV-containing target DNA may be quite small.
  • a sample of human K562 cultured cells was obtained from American Type
  • K562 is a line of pleuroeffusion cells derived
  • CML chronic myelogenous leukemia
  • the cells were grown in RPMI with 10% fetal bovine serum and maintained by
  • MDCK cells (a canine kidney cell culture) were also obtained.
  • Lysing buffer contained 0.5% SDS, 0.1 M dithiothreitol and 50 mM Tris HCL, pH 7.4. Vanadyl-ribonucleoside complex and
  • Lysing buffer was
  • RNAzol B b a Incubation time in the indicated lysing buffer.
  • b RNAzolTM (Teltest, Friends ood, TX) was used to isolate nucleic acid according to manufacturer's directions c Corresponding lane in Figure 1
  • nucleic acid precipitate was collected by centrifugation.
  • the nucleic acid pellet was recovered by resuspending in 50 ⁇ l of 10 mM Tris, 1 mM EDTA (TE), pH 7.4. A 5 ⁇ l aliquot of the
  • HeLa cells were added to 400 ⁇ l of 0.5% SDS and an equal volume (400 ⁇ l) of
  • Figure 2 shows l/20th of the resuspended nucleic acid after electrophoresis on a 0.8% agarose gel containing
  • the reaction mixture contained 20 ⁇ l of a 5X buffer (containing 200 mM Tris HCI, pH 8.1,150 mM MgCl 2 , 100 mM KCl, 50 mM dithiothreitol, 20 mM spermidine), 5 ⁇ l (15 pmol) of each
  • transcriptase 2 units of E. coli Ribonuclease H and 1000 units of T7 RNA polymerase
  • Nucleic acid in the aliquots was immobilized on the nitrocellulose by baking at 80°C.
  • the filters were prewetted with hybridization solution (6X SSC, 10X Denhardt's
  • nucleic acid isolated according to Example 1 As shown in the slot blot of Figure 3, nucleic acid isolated according to Example
  • nucleic acid was isolated as described in Example 1, except that after separating the precipitated protein, residual protein was removed from the nucleic
  • nucleic acid After removal of the protein-extracting agent, nucleic acid sequence, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids
  • acid-precipitating agents may be used in accordance with the invention.
  • the precipitated nucleic acid was resuspended in 50 ⁇ l of TE, pH 7.4, and a 5 ⁇ l
  • Figure 5 shows a photograph of the gel; the lane numbers correspond
  • Lane 1 contains Hind ]ffl-digested ⁇ DNA.
  • nucleic acid-precipitating agents such as isopropanol and CTAB are effective with either a KCl or a KOAc salt solution.
  • Table 3 Nucleic Acid Isolation Solutions
  • EXAMPLE 6 Demonstration of Substantially Intact RNA in Isolated Nucleic Acid.
  • K562 cells contain the bcr2-abl2 and the bcr3-abl2 translocations of
  • the messenger RNA produced by the bcr-abl translocation is about 8
  • the gel was then soaked in DEPC-treated water for 45 minutes and in 20X SSC for 45
  • nitrocellulose membrane After transfer, the nitrocellulose was prewetted with
  • oligonucleotide (SEQ. ID No.: 3) complementary to the junction sequence of bcr2-abl2
  • RNA recovered in accordance with the invention can be used to generate a template suitable for PCR amplification.
  • strand cDNA were: 3 ⁇ l 0.1M dithiothreitol, 50 pmol random hexamer, 4.0 ⁇ l of dNTP (2.5 mM each nucleotide), 3.0 ⁇ l 5X buffer (250 mM Tris pH 8.3, 375 mM KCl, 15 mM MgCl 2 ), 1.0 ⁇ l RNAsin (40 units/ ⁇ l), 1.0 ⁇ l superscript reverse transcriptase (200
  • the PCR reaction contained 10
  • thermocycle conditions were 35 cycles of 1 minute at 95°C, 1 minute at 55°C and 1
  • primers BB160 and BB165 is clearly visible in Figure 7, lanes 9-10.
  • primers BB160 and BB165 is clearly visible in Figure 7, lanes 9-10.
  • Nucleic acid was isolated from SiHa cells as described in Example 6. The
  • precipitated nucleic acid was resuspended in 50 ⁇ l of TE, pH 7.4, and 1, 5, and 10 ⁇ l of
  • the nucleic acid was used as a template for PCR.
  • the PCR reaction contained were 10
  • the PCR mixture was incubated for 7 minutes at 95°C and amplification initiated by
  • thermocycle conditions were the
  • MY09/MY11 amplifies a 449-458 base pair portion of the human papillomavirus (HPV)
  • LI coat gene of genital HPV strains The PCR reaction product was electrophoresed at 100 V for 1 hour on a 1.5 % agarose gel containing 1 ytg/ml of ethidium bromide.
  • nucleic acid isolated from HeLa cells in Example 2 was
  • Figure 8 is a photograph of a 1.5%
  • Cervical swab samples were obtained from a referral clinic in Boston, MA.
  • the proteinase K was denatured at 95°C for 10 minutes and a 10 ⁇ l aliquot of each sample was used as a
  • the primers used were MY09/MY11 and GH20/PCO4.
  • GH20/PCO4 amplifies a 296 bp region of the human ⁇ -globin gene and was used as a
  • precipitated material was used as a template for PCR; the attempt was unsuccessful, in
  • Example 9 Four hundred ⁇ l of 1.6 M KCl, 50 mM MgCl 2 was added
  • Nucleic acid was recovered by centrifugation at 14,000 rpm in a microfuge for 15 min. The visible pellet was allowed to dry at room temperature for one-half hour, and subsequently resuspended in 100 ⁇ l autoclaved distilled H 2 0. A 10 ⁇ l aliquot of each
  • PCR was carried out as described in Example 7, using the
  • nucleic acid was isolated according to the invention, a ⁇ -globin PCR product was visible

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Abstract

Procédé d'isolation d'ADN et d'ARN sensiblement intact à partir d'un échantillon biologique. Le procédé consiste à réaliser l'incubation de l'échantillon à l'aide d'un tampon de lyse comportant un détergent ionique et étant sensiblement exempt de composés de guanidine, à ajouter au mélange incubé une solution salée, à séparer de l'acide nucléique en solution la protéine et le détergent précipités, à ajouter à la solution d'acide nucléique un agent provoquant la précipitation de l'acide nucléique, puis à récupérer l'acide nucléique précipité. L'acide nucléique ainsi isolé est utilisable dans le clonage, dans la construction de bibliothèques et comme matrice dans les réactions d'amplification.
PCT/US1995/007940 1994-06-23 1995-06-23 Procede d'isolation rapide d'acide nucleique WO1996000228A1 (fr)

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AU29079/95A AU2907995A (en) 1994-06-23 1995-06-23 Method for the rapid isolation of nucleic acid

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US26455694A 1994-06-23 1994-06-23
US08/264,556 1994-06-23

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Cited By (32)

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WO2003070945A1 (fr) * 2002-02-15 2003-08-28 Gentr A Systems, Inc. Procede d'isolement d'adn
EP1306446A4 (fr) * 2000-08-02 2004-10-20 Oriental Yeast Co Ltd Trousses permettant d'extraire un acide nucleique et procede d'extraction d'un acide nucleique utilisant ces trousses
EP1476573A4 (fr) * 2002-01-28 2005-12-14 Ambion Inc Derives biologiques bruts capables d'une detection d'acide nucleique
WO2006089259A1 (fr) * 2005-02-18 2006-08-24 Ambion, Inc. Digestion enzymatique de tissus
WO2009014415A1 (fr) * 2007-07-23 2009-01-29 Beow Chin Yiap Procédé, kit et appareil destinés à extraire des matériels biologiques
US7964350B1 (en) 2007-05-18 2011-06-21 Applied Biosystems, Llc Sample preparation for in situ nucleic acid analysis
WO2011124703A1 (fr) * 2010-04-08 2011-10-13 Qiagen Gmbh Procédé d'isolement et de purification sélectifs d'acides nucléiques
US8211637B2 (en) 2008-12-19 2012-07-03 Life Technologies Corporation Proteinase K inhibitors, methods and compositions therefor
US8691969B2 (en) 1994-12-12 2014-04-08 Life Technologies As Isolation of nucleic acid
US20150275269A1 (en) * 2012-10-26 2015-10-01 The Emerther Company Method for purifying nucleic acid and kit
US9163228B2 (en) 2010-04-08 2015-10-20 Qiagen Gmbh Method for isolating and purifying nucleic acids
US9458494B2 (en) 2010-06-14 2016-10-04 Qiagen Gmbh Extraction of nucleic acids from wax-embedded samples
EP3464589A4 (fr) * 2016-05-31 2020-02-26 DNA Genotek Inc. Composition, système et procédé d'élimination de détergents de solutions aqueuses
US10875022B2 (en) 2007-07-13 2020-12-29 Handylab, Inc. Integrated apparatus for performing nucleic acid extraction and diagnostic testing on multiple biological samples
CN112226490A (zh) * 2020-11-12 2021-01-15 苏州创澜生物科技有限公司 一种用于dna提取的裂解缓冲液
US10900066B2 (en) 2006-03-24 2021-01-26 Handylab, Inc. Microfluidic system for amplifying and detecting polynucleotides in parallel
US10913061B2 (en) 2006-03-24 2021-02-09 Handylab, Inc. Integrated system for processing microfluidic samples, and method of using the same
US11002646B2 (en) 2011-06-19 2021-05-11 DNA Genotek, Inc. Devices, solutions and methods for sample collection
US11060082B2 (en) 2007-07-13 2021-07-13 Handy Lab, Inc. Polynucleotide capture materials, and systems using same
US11078523B2 (en) 2003-07-31 2021-08-03 Handylab, Inc. Processing particle-containing samples
US11141734B2 (en) 2006-03-24 2021-10-12 Handylab, Inc. Fluorescence detector for microfluidic diagnostic system
US11142785B2 (en) 2006-03-24 2021-10-12 Handylab, Inc. Microfluidic system for amplifying and detecting polynucleotides in parallel
US11253797B2 (en) 2010-04-08 2022-02-22 Qiagen Gmbh Chromatographic device and method for isolating and purifying nucleic acids
US11266987B2 (en) 2007-07-13 2022-03-08 Handylab, Inc. Microfluidic cartridge
US11441171B2 (en) 2004-05-03 2022-09-13 Handylab, Inc. Method for processing polynucleotide-containing samples
US11453906B2 (en) 2011-11-04 2022-09-27 Handylab, Inc. Multiplexed diagnostic detection apparatus and methods
US11466263B2 (en) 2007-07-13 2022-10-11 Handylab, Inc. Diagnostic apparatus to extract nucleic acids including a magnetic assembly and a heater assembly
US11549959B2 (en) 2007-07-13 2023-01-10 Handylab, Inc. Automated pipetting apparatus having a combined liquid pump and pipette head system
US11572581B2 (en) 2002-06-07 2023-02-07 DNA Genotek, Inc. Compositions and methods for obtaining nucleic acids from sputum
US11788127B2 (en) 2011-04-15 2023-10-17 Becton, Dickinson And Company Scanning real-time microfluidic thermocycler and methods for synchronized thermocycling and scanning optical detection
US11806718B2 (en) 2006-03-24 2023-11-07 Handylab, Inc. Fluorescence detector for microfluidic diagnostic system
US12030050B2 (en) 2006-11-14 2024-07-09 Handylab, Inc. Microfluidic cartridge and method of making same

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US4833239A (en) * 1987-01-29 1989-05-23 Genetics Institute, Inc. Method for the isolation and purification of DNA molecules
WO1989007603A1 (fr) * 1988-02-09 1989-08-24 Memorial Blood Center Of Minneapolis Isolation d'acide nucleique
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Cited By (61)

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US8691969B2 (en) 1994-12-12 2014-04-08 Life Technologies As Isolation of nucleic acid
EP1306446A4 (fr) * 2000-08-02 2004-10-20 Oriental Yeast Co Ltd Trousses permettant d'extraire un acide nucleique et procede d'extraction d'un acide nucleique utilisant ces trousses
US10011831B2 (en) 2002-01-28 2018-07-03 Applied Biosystems, Llc Crude biological derivatives competent for nucleic acid detection
US9611497B2 (en) 2002-01-28 2017-04-04 Applied Biosystems, Llc Crude biological derivatives competent for nucleic acid detection
US10233444B2 (en) 2002-01-28 2019-03-19 Applied Biosystems, Llc Crude biological derivatives competent for nucleic acid detection
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