WO2000066606A1 - Substrat contenant un detergent anionique de purification d'acide nucleique - Google Patents

Substrat contenant un detergent anionique de purification d'acide nucleique Download PDF

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Publication number
WO2000066606A1
WO2000066606A1 PCT/US2000/010225 US0010225W WO0066606A1 WO 2000066606 A1 WO2000066606 A1 WO 2000066606A1 US 0010225 W US0010225 W US 0010225W WO 0066606 A1 WO0066606 A1 WO 0066606A1
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WIPO (PCT)
Prior art keywords
substrate
nucleic acid
sds
matrix
anionic detergent
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PCT/US2000/010225
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English (en)
Inventor
Martin A. Smith
James C. Davis
Mridula Iyer
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Whatman, Inc.
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Filing date
Publication date
Application filed by Whatman, Inc. filed Critical Whatman, Inc.
Priority to EP00923408A priority Critical patent/EP1175431A4/fr
Priority to AU43535/00A priority patent/AU4353500A/en
Publication of WO2000066606A1 publication Critical patent/WO2000066606A1/fr

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    • 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

Definitions

  • the present invention relates to substrates and methods of utilizing substrates for the purification of nucleic acids or other genetic material. More specifically, the present invention relates to the purification of nucleic acids on a filter-based medium from a biological mixture of molecules in a fluid phase. Such purified nucleic acid is suitable for subsequent analysis by methods such as PCR amplification, agarose gel amplification, genotyping, sequencing, restriction digestion, and bacterial transformation.
  • Coated filter material has been shown to be useful as a matrix for the isolation and archiving of nucleic acid. Applicants have recently shown that FTA coated glass microfiber can be used as a tool for the isolation and subsequent elusion of genomic DNA. The components of the FTA coating facilitate this procedure.
  • the FTA coating comprises several components that initiate cell lysis and enable nucleic acid binding.
  • the coating also contributes to stability of the bound material.
  • the FTA coating functionally associates with the filter matrix or media.
  • the coated filter material has been shown to be useful as a tool for the rapid isolation of nucleic acid from whole cells.
  • United States Patents 5,496,562, 5,756,126, and 5,807,527 all relate to FTA coated materials for the isolation of nucleic acid from whole cells.
  • United States Patent Application 09/398,625, assigned to the assignees of the present invention discloses an FTA coated glass microfiber used as a tool for the storage, isolation, and subsequent elution of genomic DNA from whole cell samples.
  • the media employs the use of toxic choatrophic salt to initiate cell rupture and nucleic acid binding.
  • the use of a safe material and a material appropriate for obtaining cellular samples from, for example, delicate neonatal skin, and the use of a filter matrix that inherently binds nucleic acid and avoids the use of choatrophic salts to promote nucleic acid binding is desirable. Such a system would be quite useful and a significant improvement over the prior art.
  • a substrate for purifying nucleic acid consisting of a matrix and an anionic detergent fixed thereto.
  • the present invention further provides a method of preparing a filter matrix consisting of the step of fixing an anionic detergent on a matrix. Further, the present invention provides a method for isolating and archiving nucleic acid by the steps of applying a nucleic acid sample to a substrate consisting of an anionic detergent fixed to a matrix whereby the substrate physically captures the nucleic acid. The nucleic acid is then bound to the substrate.
  • FIGURE 1 shows a Polaroid image of Ameolgenin PCR products from DNA isolated from a variety of filter media and analyzed on a 1.5% agarose gel stained with ethidium bromide
  • lane 1 genomic DNA isolation carried out using GF/C glass microfiber coated with 10% SDS (medium of the invention);
  • lane 2 genomic DNA isolated using GF/C glass microfiber coated with 10% SDS/5.3M GuHC1 ;
  • lane 3 genomic DNA isolated using GF/C glass microfiber coated with FTA (FTA Elute);
  • lane 4 genomic DNA isolated using 31 -ET cellulose coated with FTA;
  • lane 5 no DNA PCR control;
  • FIGURE 2 shows a UVP Gel Documentation image of an agarose gel showing the presence of isolated plasmid DNA from bacterial colonies using a variety of coated filter materials; the three-band pattern of classic plasmid isolation being noted (supercoiled, nicked, and linear), lanes 1 and 2: Plasmid isolated using FTA coated GF/C glass microfiber (FTA Elute); lanes 3 and 4: Plasmid isolated using 10% SDS coated DBS 1000 glass microfiber; lanes 5 and 6: Plasmid isolated using FTA coated glass flake impregnated glass microfiber;
  • FIGURE 3 shows a UVP Gel Documentation image of an agase gel showing the presence of isolated plasmid DNA from liquid culture using a variety of coated filter materials in the GenSpin format; lane 1 : 500ng commercially available pGEM plasmid DNA (Promega); lanes 2-5: Plasmid isolated using FTA coated GF/C glass microfiber (FTA Elute); lanes 6-9: Plasmid isolated using 10% SDS coated DBS1000 glass microfiber; and
  • FIGURE 4 shows a UVP Gel Documentation image of an agarose gel showing the presence of different isolated plasmid DNA populations from liquid culture using a variety of SDS coated and FTA coated filter materials in the 96 well multiwell format; lane 1 : 1% SDS coated DBS1000 presented with pGEM plasmid culture; lane 2: 2.5% SDS coated DBS1000 presented with pGEM plasmid culture; lane 3: 5% SDS coated DBS1000 presented with pGEM plasmid culture; lane 4: 7.5% SDS coated DBS1000 presented with pGEM plasmid culture; lane 5: 10% SDS coated DBS1000 presented with pGEM plasmid culture; lane 6: FTA coated GF/C presented with pGEM plasmid culture; lane 7: 1% SDS coated DBS1000 presented with pUC19 plasmid culture; lane 8: 2.5% SDS coated DBS1000 presented with pUC19 plasmid culture; lane 9: 5% S
  • the present invention provides a substrate for purifying nucleic acid, the substrate consisting of a matrix and an anionic detergent affixed thereto.
  • the source of the nucleic acid can be a biological sample containing whole cells.
  • the whole cell can be, but not restricted to, blood, bacterial culture, bacterial colonies, saliva, urine, drinking water, plasma, stool samples, and sputum.
  • the samples can be collected by various means known in the art, transported to the substrate, and then applied thereto.
  • the substrate can be in the form of a sampling device, such as a swab, sheet material, or the like and the sample can be obtained directly from the source.
  • the substrate can be in the form of a device which can swipe or otherwise obtain the cell sample from a source.
  • the source can be a sample tube containing a liquid sample, an organ, such as a mouth, ear, or other part of a human or animal, a sample pool, such as a blood sample at a crime scene or the like, or other various sources of cells known in the scientific, forensic, and other arts.
  • the substrate consists of a matrix and an anionic detergent affixed thereto.
  • the anionic detergent can be selected from the group including sodium dodecyl sulfate (SDS). SDS can be obtained in various forms, such as the C 12 form and the lauryl sulfate. Other anionic detergents can be used, such as alkyl aryl sulphonates, long chain (fatty alcohol sulphates) olefine sulphates, sulphosuccinates, phosphate ester, sodium tetradecylsulphate and sodium 2-ethylhexysulphate.
  • 5%-10% SDS can be used in accordance with the present invention.
  • a definite optimum SDS concentration is achieved in the 5 - 7.5% SDS concentration range for coating particular glass microfiber in order to enrich for and purify different plasmid populations directly from liquid cultures in a multi-well format, such formats being well known in the art.
  • the matrix can be associated with the anionic detergent in different forms, well known in the coating art.
  • the anionic detergent can impregnate the matrix, such methods of manufacturing to cause impregnation being well known in the art.
  • the anionic detergent can merely coat the matrix, such coating being obtained by methods well known in the art.
  • the matrix can be made from fibers well known in the filtration art.
  • the matrix can include fibers selected from the group including cellulose, glass fibers, glass microfibers, poly(vinylidene fluoride) (PDVF), non-woven polyesters, and melt blown polypropylenes.
  • the anionic detergent, such as the SDS can be applied to the filter matrix at varying concentrations.
  • the fibrous filter matrix of the present invention can be manufactured in various forms.
  • the fibrous filter matrix can be manufactured in a sheet form, which allows for it to be in various formats such as multi-well plates, spin tubes, slides, cartridges, swabs, and pads.
  • substrate or matrix most generally means a porous material or filter media formed, but not limited to, either fully or partly from the materials set forth above, and/or their derivatives.
  • the matrix material does not inhibit the sorption of chemical coating solution and does not inhibit the storage, elusion, and subsequent analysis of nucleic acid containing material added to it. It is preferred that the matrix be of a porous nature to facilitate immobilization of nucleic acid.
  • the substrate of the present invention allows for elusion of the genetic material therefrom in a state that allows for subsequent analysis. Such elusion is a time efficient step thereby providing for almost immediately analysis.
  • the anionic detergent By being fixed to the matrix, the anionic detergent is disposed, sorbed, or otherwise associated with the matrix of the present invention such that the matrix and anionic detergent coating function together to immobilize nucleic acid thereon through an action of cellular lysis of cells presented to the support. That is, the anionic detergent can be adsorbed, absorbed, coated over, or otherwise disposed, in functional relationship with the matrix.
  • the substrate of the present invention is capable of releasing the genetic material immobilized thereon to by a heat elusion step.
  • a heat elusion is accomplished by the exposure of the support having the genetic material stored thereon to heated water, the water being nuclease-free. This capacity to allow for elusion characterizes the various support materials of the present invention.
  • the matrix of the present invention is such that nucleic acid immobilized to it can remain so in a stable form and not exhibit degradation, shearing, endonuclease digestion, or UV damage. Accordingly, the substrate in the present invention can be used to transport and archive nucleic acid samples.
  • the present invention is capable of immobilizing nucleic acid that is collected in the form of a soluble fraction followed a simplified elusion process during which immobilized nucleic acid is released from the matrix of the invention.
  • the matrix of the invention yields nucleic acid of sufficient quality that it does not impair downstream analysis, such as polymerase chain reaction (PCR), ligase chain reaction (LCR), transcripted mediated amplification (TMA), reverse transcriptase initiated PCR, DNA or RNA hybridization techniques, sequencing, and the like.
  • PCR polymerase chain reaction
  • LCR ligase chain reaction
  • TMA transcripted mediated amplification
  • reverse transcriptase initiated PCR DNA or RNA hybridization techniques, sequencing, and the like.
  • Nucleic acid immobilization to a solid support although a suitable template for singular PCR reactions, cannot be measured or detected by traditional techniques such as optical density or fluorescence. Nucleic acid must be in solution for these techniques.
  • Other post-purification techniques wherein nucleic acid is desired in the soluble form includes cloning, hybridization protection assay, bacterial transformation, mammalian transfection, transcription-mediated amplification, and other such methods.
  • the present invention provides nucleic acid in such a soluble form.
  • the present inventive substrate can be prepared by fixing an anionic detergent on a matrix. The fixing step can be achieved by either impregnating the anionic detergent within the matrix or coating the matrix, as discussed above.
  • the present invention provides a method for isolating and archiving nucleic acid by the general steps of applying a nucleic acid sample to a substrate consisting of the anionic detergent fixed to the matrix, the substrate physically capturing the nucleic acid, and then bonding the nucleic acid to the substrate.
  • the bonding step is achieved by heating the substrate having nucleic acid applied thereto, by the method discussed above.
  • the applying step can be achieved by applying whole cells to the substrate.
  • the substrate itself actually induces the lysing of the cells thereby releasing the nucleic acid into the substrate.
  • the substrate presents a vast surface area upon which the nucleic acid is bound.
  • a washing step such as with various buffers set forth in the example section, but not limited thereto, can be achieved and is done after cell lysis.
  • the substrate then physically captures the nucleic acid within the intrastaces thereof.
  • the bound nucleic acid can be released from the substrate for further processing and analysis.
  • the release is achieved by washing steps at elevated temperature, as demonstrated in the examples below.
  • enrichment for different populations of nucleic acid from the same cell source can be achieved using incremental temperature regimes.
  • plasmid DNA can be isolated and enriched from bacterial colonies using the substrate of the present invention.
  • Populations, such as larger populations of supercoiled plasmid, followed by nicked plasmid and finally by linear plasmid migrating to the top of the isolating gel can be achieved utilizing incremental increases in incubation temperature.
  • the FTA coating cocktail of the prior art contains 2% SDS. It is unlikely that this percentage can be increased due to saturation points when in conjunction with other components of the cocktail. This limits the lysing capability of the FTA coating filters of the prior art as a critical micelle concentration of SDS can be easily reached when presented with large numbers of cells, such as with a bacterial colony. Therefore, substrates containing a greater concentration of the lytic agent, the anionic detergent, enable greater lysing capability and in turn, greater nucleic acid recoveries. This is demonstrated in the examples set forth below.
  • the following examples demonstrate the preparation, function, and utility of the present invention in order to isolate nucleic acid from various sources.
  • the examples further demonstrate the effect of SDS coating concentrations on glass microfiber for the purification of direct plasmids from direct liquid culture using a multiwell filter plate.
  • the following examples demonstrate that the present invention can be used for the rapid purification and isolation of nucleic acids and genetic material from samples containing whole cells.
  • Example 1 The Use of 10% SDS Coated Glass Microfiber For The Purification Of Genomic DNA From Whole Blood Samples:
  • GF/C glass microfiber was cut to size and pieces soaked in a dish for one hour at room temperature in a solution of either 10% SDS (filter matrix of the invention) or 10% SDS/5.3M GuHC1. After soaking the wetted filters were placed in a convection oven and heated for thirty minutes at 80°C. This soaking and baking process constitutes the coating process. To the coated filter materials, as well as FTA coated GF/C and FTA coated 31 -ET cellulose, several drops of freshly finger stick drawn blood was spotted and allowed to air dry for two minutes. Once dried a 1 mm punch was taken from the dried blood spots and applied to individual 200ul polypropylene PCR tubes.
  • FTA Purification Reagent (0.29g NaCI, 5ml 1M Tris pH 7.5, 1ml 0.5M EDTA, 2.5ml Triton X-100, per 500ml) (Fitzco, Inc.) was added. Tubes were incubated for five minutes at room temperature with no shaking. Following incubation of the FTA Purification Reagent was aspirated from the tube. A second aliquot of 200ul of FTA Purification Reagent was added to each tube. The tubes were incubated for five minutes at room temperature without shaking. Following incubation, the FTA Purifcation Reagent was aspirated from all the tubes.
  • TE 10mM Tris-HCI, 0.1mM EDTA, pH 8.0
  • 20ul of nuclease free water was then applied to every tube.
  • Each tube was then administered to a thermal cycler and heated to 95°C for ten minutes. Following heat incubation the 20ul of water was removed and dispensed into individual 25ul Amelogenin PCR reactions.
  • PCR amplification was carried out following parameters described by the manufacturer of the Amelogenin primer set (Promega). Following PCR 10ul of each PCR reaction was visualized on a 1.5% agarose gel stained with ethidium bromide, and photographed using a Polaroid camera.
  • Lane 1 is GF/C glass microfiber coated with 10% SDS (filter media of this invention) and lane 3 is FTA coated glass microfiber.
  • 10% SDS coated GF/C functions in terms of lysis, binding, and elution as well as the FTA coated GF/C.
  • the advantage is that one does not have to manufacture the 10% SDS GF/C with the same level of control as the FTA coated GF/C which has multiple components within the FTA cocktail.
  • PCR-ready genomic DNA has not been released from glass microfiber coated with a mixture of 10% SDS and 5.3M GuHC1 (lane 2), and FTA coated 31-ET cellulose (lane 4).
  • the non-appearance of PCR product in lanes 3 and 4 may be due to the inability of the filter material and coating to lyse cells. More probably, this effect is caused by an inability of the filter material and coating to release the bound genomic DNA after lysing (U.S. Patents 5,496,562, 5,756,126, 5,807,527).
  • Lane 5 is a no DNA PCR control.
  • DH5 ⁇ bacterial culture was transformed with the plasmid pGEM and streaked onto an LB medium petri dish. Following overnight incubation of the streaked petri dish at 37°C, individual bacterial colonies were picked and resuspended in 10ul of PBS buffer. The resuspended colonies were then spotted to 7mm discs of either FTA coated GF/C glass microfiber, 10% SDS coated DBS1000 glass microfiber (the medium of the invention), and FTA coated glass flake impregnated glass microfiber. After spotting, the discs were placed into a 2ml Eppendorf tube.
  • TE buffer containing 10ug/ml Rnase was added and incubated for five minutes at room temperature without shaking. After incubation, the TE/Rnase buffer was aspirated from the tubes and replaced with 1ml of TE buffer an dincubated for five minutes at room temperature without shaking. Again, TE was aspirated and replaced with fresh TE buffer following incubation. Following the final TE buffer incubation, 200ul of nuclease free water was added to each disc and incubated at 65°C for fifteen minutes. Applicants have previously shown (Iyer, M. et al. (2000) Rapid Plasmid Template Preparation using Novel Paper Based Systems. Abstract & Poster.
  • plasmid DNA can be isolated and enriched for from bacterial colonies using coated filter media.
  • the classic three-band appearance of plasmic on the gel is noted (larger populations of supercoiled plasmid, followed by nicked plasmid and finally linear plasmid migrating at the top of the tel).
  • FTA coating cocktail contains 2% SDS, and that this percentage is unlikely to be increased due to saturation points when in conjunction with other components of the cocktail.
  • TE buffer containing 10ug/ml of Rnase was added and incubated for one minute at room temperature before centrifugation at 12,000 xg for one minute.
  • the TE step was repeated twice more (without Rnase).
  • 200ul of nuclease free water was added to the spin basket of each set of GenSpin tubes. Tubes were then incubated at 55°C for fifteen minutes in a water bath. Following heat incubation, the tubes were subjected to a final centrifugation step at 12,000 xg for two minutes. 20ul of the final filtrate was analyzed by agarose gel electrophoresis. The ethidium bromide stain agarose gel image was captured using UVP Gel Documentation System.
  • SDS coated filter material in the GenSpin format, Applicants have been able to purify high quality plasmid DNA directly from liquid culture without the need for cell harvesting, lysis, precipitation and binding that is required for traditional chaotrophic salt plasmid isolations. Furthermore, the SDS coated material is more effective at purifying plasmid directly from liquid culture than FTA coated filter material.
  • Example 4 Effect Of SDS Coating Concentration On Glass Microfiber For the Purification Of Different Plasmids From Direct Liquid Culture Using A Multiwell Filterplate
  • DH5 ⁇ bacterial cells were transformed with either pGEM, pUC19, or pGEM.SPORT plasmids and grown overnight in separate 10ml LB cultures at 37°C with constant shaking. To each well of the filled multiwell plate, a 500ul aliquot of the bacterial culture was added.
  • the multiwell plate was placed on top of a multiwell collection plate and centrifuged at 1000 xg for two minutes so as to transfer the nuclease free water within the filter multiwell plate to the collection plate. 20ul of the final filtrate was analyzed by agarose gel electrophoresis. The ethidium bromide stained agarose gel image was captured using UVP Gel Documentation System.

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Abstract

L'invention a trait à un substrat servant à purifier un acide nucléique et à enrichir des populations d'acides nucléiques à partir d'une seule source de cellule, et comprenant une matrice et un détergent anionique fixé sur cette dernière. Un procédé de fabrication du substrat consiste à attacher un détergent anionique à la matrice. Un procédé d'isolation et d'archivage d'acide nucléique utilisant la matrice consiste à appliquer l'échantillon d'acide nucléique sur le substrat, ce dernier capturant physiquement l'acide nucléique et le liant au substrat.
PCT/US2000/010225 1999-04-30 2000-04-14 Substrat contenant un detergent anionique de purification d'acide nucleique WO2000066606A1 (fr)

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EP00923408A EP1175431A4 (fr) 1999-04-30 2000-04-14 Substrat contenant un detergent anionique de purification d'acide nucleique
AU43535/00A AU4353500A (en) 1999-04-30 2000-04-14 Substrate including anionic detergent for purifying nucleic acid

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US13190999P 1999-04-30 1999-04-30
US60/131,909 1999-04-30

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Cited By (15)

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WO2003106031A1 (fr) * 2002-06-12 2003-12-24 F. Hoffmann-La Roche Ag Procede et dispositif de detection de la presence d'un analyte dans un echantillon pour essai
EP1436405A2 (fr) * 2001-08-20 2004-07-14 Whatman Inc. Purification et recuperation d'adn a partir d'echantillons a haute teneur en particules et en solides
EP1451320A2 (fr) * 2001-11-15 2004-09-01 Whatman Inc. Materiaux et methodes pour liberation de materiau genetique
EP1453484A2 (fr) * 2001-11-15 2004-09-08 Whatman, Inc. Methodes et materiels de detection de materiel genetique
JP2013515494A (ja) * 2009-12-29 2013-05-09 ワットマン インコーポレイテッド 核酸溶出の改良
US9040675B2 (en) 2012-04-30 2015-05-26 General Electric Company Formulations for nucleic acid stabilization on solid substrates
US9040679B2 (en) 2012-04-30 2015-05-26 General Electric Company Methods and compositions for extraction and storage of nucleic acids
US9044738B2 (en) 2012-04-30 2015-06-02 General Electric Company Methods and compositions for extraction and storage of nucleic acids
CN105907751A (zh) * 2016-06-27 2016-08-31 河南农业大学 基于fta卡从粪便中提取隐孢子虫卵囊dna的方法
US9480966B2 (en) 2012-04-30 2016-11-01 General Electric Company Substrates and methods for collection, stabilization and elution of biomolecules
US10435735B2 (en) 2014-03-07 2019-10-08 Dna Genotek Inc. Composition and method for stabilizing nucleic acids in biological samples
US10638963B2 (en) 2017-01-10 2020-05-05 Drawbridge Health, Inc. Devices, systems, and methods for sample collection
US11002646B2 (en) 2011-06-19 2021-05-11 DNA Genotek, Inc. Devices, solutions and methods for sample collection
US11266337B2 (en) 2015-09-09 2022-03-08 Drawbridge Health, Inc. Systems, methods, and devices for sample collection, stabilization and preservation
US11572581B2 (en) 2002-06-07 2023-02-07 DNA Genotek, Inc. Compositions and methods for obtaining nucleic acids from sputum

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EP1436405A4 (fr) * 2001-08-20 2006-01-04 Whatman Inc Purification et recuperation d'adn a partir d'echantillons a haute teneur en particules et en solides
EP1436405A2 (fr) * 2001-08-20 2004-07-14 Whatman Inc. Purification et recuperation d'adn a partir d'echantillons a haute teneur en particules et en solides
US6972329B2 (en) 2001-11-15 2005-12-06 Whatman, Inc. Materials and methods for releasing genetic material
EP1453484A2 (fr) * 2001-11-15 2004-09-08 Whatman, Inc. Methodes et materiels de detection de materiel genetique
EP1451320A4 (fr) * 2001-11-15 2005-03-02 Whatman Inc Materiaux et methodes pour liberation de materiau genetique
EP1453484A4 (fr) * 2001-11-15 2005-03-09 Whatman Inc Methodes et materiels de detection de materiel genetique
EP1451320A2 (fr) * 2001-11-15 2004-09-01 Whatman Inc. Materiaux et methodes pour liberation de materiau genetique
US7504363B2 (en) 2001-11-15 2009-03-17 Whatman, Inc. Methods using four-layer filter for PCR sample preparation
US11572581B2 (en) 2002-06-07 2023-02-07 DNA Genotek, Inc. Compositions and methods for obtaining nucleic acids from sputum
WO2003106031A1 (fr) * 2002-06-12 2003-12-24 F. Hoffmann-La Roche Ag Procede et dispositif de detection de la presence d'un analyte dans un echantillon pour essai
JP2013515494A (ja) * 2009-12-29 2013-05-09 ワットマン インコーポレイテッド 核酸溶出の改良
US11002646B2 (en) 2011-06-19 2021-05-11 DNA Genotek, Inc. Devices, solutions and methods for sample collection
US11592368B2 (en) 2011-06-19 2023-02-28 DNA Genotek, Inc. Method for collecting and preserving a biological sample
US11549870B2 (en) 2011-06-19 2023-01-10 DNA Genotek, Inc. Cell preserving solution
US11536632B2 (en) 2011-06-19 2022-12-27 DNA Genotek, Inc. Biological collection system
US9480966B2 (en) 2012-04-30 2016-11-01 General Electric Company Substrates and methods for collection, stabilization and elution of biomolecules
US10625242B2 (en) 2012-04-30 2020-04-21 General Electric Company Substrates and methods for collection, stabilization and elution of biomolecules
US9044738B2 (en) 2012-04-30 2015-06-02 General Electric Company Methods and compositions for extraction and storage of nucleic acids
US9040679B2 (en) 2012-04-30 2015-05-26 General Electric Company Methods and compositions for extraction and storage of nucleic acids
US9040675B2 (en) 2012-04-30 2015-05-26 General Electric Company Formulations for nucleic acid stabilization on solid substrates
US10435735B2 (en) 2014-03-07 2019-10-08 Dna Genotek Inc. Composition and method for stabilizing nucleic acids in biological samples
US11198899B2 (en) 2014-03-07 2021-12-14 Dna Genotek Inc. Composition and method for stabilizing nucleic acids in biological samples
US11266337B2 (en) 2015-09-09 2022-03-08 Drawbridge Health, Inc. Systems, methods, and devices for sample collection, stabilization and preservation
CN105907751A (zh) * 2016-06-27 2016-08-31 河南农业大学 基于fta卡从粪便中提取隐孢子虫卵囊dna的方法
US10638963B2 (en) 2017-01-10 2020-05-05 Drawbridge Health, Inc. Devices, systems, and methods for sample collection
US10888259B2 (en) 2017-01-10 2021-01-12 Drawbridge Health, Inc. Cartridge assemblies for storing biological samples
US10932710B2 (en) 2017-01-10 2021-03-02 Drawbridge Health, Inc. Carriers for storage and transport of biological samples
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