US20120289690A1 - Nucleic acid elution - Google Patents

Nucleic acid elution Download PDF

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Publication number
US20120289690A1
US20120289690A1 US13/519,391 US201013519391A US2012289690A1 US 20120289690 A1 US20120289690 A1 US 20120289690A1 US 201013519391 A US201013519391 A US 201013519391A US 2012289690 A1 US2012289690 A1 US 2012289690A1
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Prior art keywords
dna
solid matrix
matrix
inulin
peg
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Abandoned
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US13/519,391
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English (en)
Inventor
Andrew Francis Page
Breck Olland Parker
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Global Life Sciences Solutions USA LLC
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GE Healthcare Bio Sciences Corp
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Priority to US13/519,391 priority Critical patent/US20120289690A1/en
Assigned to WHATMAN, INC. reassignment WHATMAN, INC. ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: PAGE, ANDREW FRANCIS, PARKER, BRECK OLLAND
Assigned to GE HEALTHCARE BIO-SCIENCES CORP. reassignment GE HEALTHCARE BIO-SCIENCES CORP. MERGER (SEE DOCUMENT FOR DETAILS). Assignors: WHATMAN, INC.
Publication of US20120289690A1 publication Critical patent/US20120289690A1/en
Assigned to GLOBAL LIFE SCIENCES SOLUTIONS USA LLC reassignment GLOBAL LIFE SCIENCES SOLUTIONS USA LLC CHANGE OF NAME (SEE DOCUMENT FOR DETAILS). Assignors: GE HEALTHCARE BIO-SCIENCES CORP.
Abandoned legal-status Critical Current

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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
    • 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

Definitions

  • This invention relates to the storage on a solid matrix of genetic material, in particular DNA that has been purified prior to the application to the solid matrix.
  • Burgoyne (WO 90/03959) described a method whereby biological samples, usually blood, could be applied to a solid matrix which combined reagents which lysed the cells. The released DNA was retained on the solid matrix. These samples could be stored for long periods at room temperature.
  • inulin a polysaccharide found in plants
  • FIG. 1 shows the structure of inulin.
  • PEG is prepared in a variety of molecuar weights defined by the average number of subunits per molecule. Polymers of MW 400, 1000 and 3350 were evaluated. It was observed that PEG 1000 produced the best recovery of applied DNA results (data not shown) and was used in the remainder of the tests but is referred to as PEG. At concentration of PEG at about 25% the results varied between experiments; this may imply small inconsistencies in the coating process of the solid matrix at these concentrations.
  • Inulin is a naturally occurring polysaccharide found in many plants. Its structure is given in FIG. 1 . it is anticipated that simple modifications of inulin eg esterification would be possible and still achieve the improved elution of the applied purified DNA.
  • the purified DNA can be applied to the solid matrix that has been treated with inulin in buffers that are routinely used in nucleic acid chemistry. Up to 10% PEG can also be included in the application buffer.
  • the DNA prior to application to the solid matrix can be purified by a variety of standard laboratory techniques.
  • the solid matrix was FTA Elute 903 matrix from Whatman.
  • a 4 M stock of guanidine thiocyanate was prepared and diluted to 2 M using various concentrations of test reagents.
  • 903 matrix (2 ⁇ 2 1 ⁇ 4′′) was placed into trays containing guanidine thiocyanate/test reagent mixes and agitated gently for 10 seconds.
  • Matrices were dried for 10 min on a metal rack using two hair dryers (Simply Basic DS-727); one placed at a 30° angle 15 cm above the matrix, the other placed 25 cm below the matrix at a 30° angle such that the two hair dryers and the matrix were in alignment. Matrices were dried further without the air flow at 21 ⁇ 2° C. overnight. 4) Matrices were stored at room temperature in a desiccator until use.
  • Human DNA (Roche) was spotted onto the test solid matrix at concentration of 160 ng/ ⁇ l. Usually a pre-punched 5 mm diameter disc of the matrix was used. Discs were dried at room temperature for a minimum of 3 hours.
  • Quantification of DNA in eluates was performed by QPCR using a 7900HT Thermal Cycler (Applied Biosystems). Reactions were set up using an RNase P assay and TAQMAN® Universal PCR Master Mix (Applied Biosystems). A four point standard curve was prepared using a serial dilution from 10 to 0.01 ng/ ⁇ l of the same Roche DNA used for experiments. Early QPCR quantifications were performed in 96-well plates and were set up manually. Following the introduction and validation of a liquid handling robot, later quantifications were performed using 384-well plates. Both 96 and 384-well plates were validated and also tested against each other to check for consistency between methods.
  • Results show that matrices impregnated with either inulin or PEG did 20 result in increased DNA recovery compared to FTA Elute by itself.
  • the use of a spotting buffer containing 10% PEG resulted in an additional increase in DNA yield (85% when used with FTA Elute+20% inulin).
  • the matrix containing 20% inulin showed the highest % recovery of applied purified DNA. Since PEG was also identified as a possible additive to matrix impregnation chemistry, FTA Elute impregnated with a combination of 20% inulin and 10% PEG was also prepared for further investigation. A spotting buffer containing 10% PEG was confirmed as further increasing yields when used in conjunction with the modified matrix chemistry.
  • Results were also obtained from experiments where the discs with applied purified DNA had been stored in a dessicator at room temperature. The results showed that the discs could be stored for at least twenty-three days before DNA elution with similar increased recovery of DNA when the matrix had been treated with inulin.
  • Results also showed that the amount of DNA applied and recovered from the test matrix could be as low as 1 ng and as high as 1 ⁇ g (the maximum tested) and the increased effect on inulin treatment was still observed.

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  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Genetics & Genomics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • Analytical Chemistry (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • Microbiology (AREA)
  • General Health & Medical Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Biophysics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Immunology (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Plant Pathology (AREA)
  • Saccharide Compounds (AREA)
  • Polysaccharides And Polysaccharide Derivatives (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Medicinal Preparation (AREA)
US13/519,391 2009-12-29 2010-12-21 Nucleic acid elution Abandoned US20120289690A1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
US13/519,391 US20120289690A1 (en) 2009-12-29 2010-12-21 Nucleic acid elution

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US29065209P 2009-12-29 2009-12-29
US13/519,391 US20120289690A1 (en) 2009-12-29 2010-12-21 Nucleic acid elution
PCT/EP2010/070389 WO2011080160A1 (en) 2009-12-29 2010-12-21 Improvements to nucleic acid elution

Related Parent Applications (1)

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PCT/EP2010/070389 A-371-Of-International WO2011080160A1 (en) 2009-12-29 2010-12-21 Improvements to nucleic acid elution

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US17/122,606 Division US11725201B2 (en) 2009-12-29 2020-12-15 Nucleic acid elution

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US20120289690A1 true US20120289690A1 (en) 2012-11-15

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US13/519,391 Abandoned US20120289690A1 (en) 2009-12-29 2010-12-21 Nucleic acid elution
US17/122,606 Active 2031-08-03 US11725201B2 (en) 2009-12-29 2020-12-15 Nucleic acid elution
US18/340,609 Active US12247199B2 (en) 2009-12-29 2023-06-23 Nucleic acid elution

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US18/340,609 Active US12247199B2 (en) 2009-12-29 2023-06-23 Nucleic acid elution

Country Status (7)

Country Link
US (3) US20120289690A1 (https=)
EP (1) EP2519631B1 (https=)
JP (1) JP5899118B2 (https=)
CN (1) CN102725406B (https=)
AU (1) AU2010338349B2 (https=)
CA (1) CA2785913C (https=)
WO (1) WO2011080160A1 (https=)

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9534214B2 (en) 2013-10-31 2017-01-03 General Electric Company Substrates and associated methods for elution of nucleic acids
US9719082B2 (en) 2013-10-31 2017-08-01 General Electric Company Substrates and associated methods for elution of nucleic acids
US10625242B2 (en) 2012-04-30 2020-04-21 General Electric Company Substrates and methods for collection, stabilization and elution of biomolecules
US10638963B2 (en) 2017-01-10 2020-05-05 Drawbridge Health, Inc. Devices, systems, 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
US11795446B2 (en) 2020-03-23 2023-10-24 Ricoh Company, Ltd. Carrier and testing method

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2519631B1 (en) * 2009-12-29 2017-02-15 GE Healthcare Bio-Sciences Corp. Improvements to nucleic acid elution
US10000742B2 (en) * 2015-11-19 2018-06-19 General Electric Company Device and method of collection for RNA viruses
CN105462961B (zh) * 2016-01-14 2019-04-19 苏州市公安局 一种提取核酸的方法及试剂盒

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5124444A (en) * 1989-07-24 1992-06-23 Microprobe Corporation Lactam-containing compositions and methods useful for the extraction of nucleic acids
WO2003044211A2 (en) * 2001-11-15 2003-05-30 Whatman Inc. Materials and methods for releasing genetic material
US20060147944A1 (en) * 2005-01-04 2006-07-06 Piotr Chomczynski Reagents and methods for storage and processing of biological samples for DNA analysis
US20060153920A1 (en) * 2004-09-24 2006-07-13 Ketan Amin Lyophilized pharmaceutical compositions

Family Cites Families (12)

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WO1990003959A1 (en) 1988-10-05 1990-04-19 The Flinders University Of South Australia Solid medium and method for dna storage
US5496562A (en) * 1988-10-05 1996-03-05 Flinders Technologies Pty Ltd Solid medium and method for DNA storage
GB8903593D0 (en) * 1989-02-16 1989-04-05 Pafra Ltd Storage of materials
US5939259A (en) 1997-04-09 1999-08-17 Schleicher & Schuell, Inc. Methods and devices for collecting and storing clinical samples for genetic analysis
AU4353500A (en) * 1999-04-30 2000-11-17 Whatman, Inc. Substrate including anionic detergent for purifying nucleic acid
SE9904475D0 (sv) * 1999-12-08 1999-12-08 Artursson Nucleic acid delivery system
JP3776334B2 (ja) * 2001-07-11 2006-05-17 株式会社富士薬品 イヌリン含有製剤
CA2484062A1 (en) * 2001-08-20 2003-02-27 James C. Davis Dna purification and recovery from high particulate and solids samples
CA2465961A1 (en) * 2001-11-06 2003-05-15 Cortex Biochem, Inc. Isolation and purification of nucleic acids
US20040033546A1 (en) * 2002-04-10 2004-02-19 The Trustees Of Columbia University In The City Of New York Novel microarrays and methods of use thereof
BRPI0415850A (pt) * 2003-10-23 2007-01-02 Alza Corp composição de dna estabilizado para o revestimento de micro projeções
EP2519631B1 (en) * 2009-12-29 2017-02-15 GE Healthcare Bio-Sciences Corp. Improvements to nucleic acid elution

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5124444A (en) * 1989-07-24 1992-06-23 Microprobe Corporation Lactam-containing compositions and methods useful for the extraction of nucleic acids
WO2003044211A2 (en) * 2001-11-15 2003-05-30 Whatman Inc. Materials and methods for releasing genetic material
US20060153920A1 (en) * 2004-09-24 2006-07-13 Ketan Amin Lyophilized pharmaceutical compositions
US20060147944A1 (en) * 2005-01-04 2006-07-06 Piotr Chomczynski Reagents and methods for storage and processing of biological samples for DNA analysis

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
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de Jonge et al. (European Journal of Pharmaceutical Sciences (2007), 32(1), 33-44) *
Kravchenko et al. (Russian Journal of Bioorganic Chemistry, 2006, Vol. 32, No. 6, pp. 547–551). *

Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10625242B2 (en) 2012-04-30 2020-04-21 General Electric Company Substrates and methods for collection, stabilization and elution of biomolecules
US9534214B2 (en) 2013-10-31 2017-01-03 General Electric Company Substrates and associated methods for elution of nucleic acids
US9719082B2 (en) 2013-10-31 2017-08-01 General Electric Company Substrates and associated methods for elution of nucleic acids
US11266337B2 (en) 2015-09-09 2022-03-08 Drawbridge Health, Inc. Systems, methods, and devices for sample collection, stabilization and preservation
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
US11298060B2 (en) 2017-01-10 2022-04-12 Drawbridge Health, Inc. Devices for collecting biological samples
US11795446B2 (en) 2020-03-23 2023-10-24 Ricoh Company, Ltd. Carrier and testing method

Also Published As

Publication number Publication date
CN102725406B (zh) 2016-06-01
CA2785913A1 (en) 2011-07-07
AU2010338349A1 (en) 2012-06-21
EP2519631B1 (en) 2017-02-15
US20210102190A1 (en) 2021-04-08
JP5899118B2 (ja) 2016-04-06
US11725201B2 (en) 2023-08-15
AU2010338349B2 (en) 2015-03-05
CN102725406A (zh) 2012-10-10
JP2013515494A (ja) 2013-05-09
WO2011080160A1 (en) 2011-07-07
EP2519631A1 (en) 2012-11-07
US20230332133A1 (en) 2023-10-19
US12247199B2 (en) 2025-03-11
CA2785913C (en) 2019-05-21

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