CA2319691A1 - Eluting reagents, methods and kits for isolating dna - Google Patents

Eluting reagents, methods and kits for isolating dna Download PDF

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Publication number
CA2319691A1
CA2319691A1 CA002319691A CA2319691A CA2319691A1 CA 2319691 A1 CA2319691 A1 CA 2319691A1 CA 002319691 A CA002319691 A CA 002319691A CA 2319691 A CA2319691 A CA 2319691A CA 2319691 A1 CA2319691 A1 CA 2319691A1
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Canada
Prior art keywords
dna
reagent
eluting reagent
dna eluting
total volume
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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.)
Granted
Application number
CA002319691A
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French (fr)
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CA2319691C (en
Inventor
Ellen M. Heath
Ruth M. Shuman
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.)
Qiagen North American Holdings Inc
Original Assignee
Gentra Systems, Inc.
Ellen M. Heath
Ruth M. Shuman
Qiagen North American Holdings, Inc.
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Application filed by Gentra Systems, Inc., Ellen M. Heath, Ruth M. Shuman, Qiagen North American Holdings, Inc. filed Critical Gentra Systems, Inc.
Publication of CA2319691A1 publication Critical patent/CA2319691A1/en
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Publication of CA2319691C publication Critical patent/CA2319691C/en
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    • 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
    • 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
    • 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

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  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Organic Chemistry (AREA)
  • Biomedical Technology (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Analytical Chemistry (AREA)
  • Molecular Biology (AREA)
  • General Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Biochemistry (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Plant Pathology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Immunology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Saccharide Compounds (AREA)

Abstract

Eluting reagents and methods for isolating DNA from biological materials are provided.

Claims (38)

1. A DNA eluting reagent comprising:
(a)~a buffer;
(b)~a base;
(c)~a chelating agent; and (d)~water.
2. The DNA eluting reagent of claim 1, wherein the chelating agent is used in an amount no greater than about 0.1 mM, based on the total volume of the DNA eluting reagent.
3. The DNA eluting reagent of claim 1, wherein the DNA eluting reagent has a pH of at least about 7.
4. The DNA eluting reagent of claim 1, wherein the base is chosen from the group consisting of alkaline metal hydroxides.
5. The DNA eluting reagent of claim 1, wherein the buffer is present in an amount no greater than 20 mM, based on the total volume of the DNA eluting reagent.
6. The DNA eluting reagent of claim 1, wherein the buffer is present in an amount from 0.001-20 mM, based on the total volume of the DNA eluting reagent.
7. The DNA eluting reagent of claim 1, wherein the buffer is present in an amount from 0.01-15 mM, based on the total volume of the DNA eluting reagent.
8. The DNA eluting reagent of claim 1, wherein the base is present in an amount no greater than 20 mM, based on the total volume of the DNA eluting reagent.
9. The DNA eluting reagent of claim 1, wherein the base is present in an amount from 1-10 mM, based on the total volume of the DNA eluting reagent.
10. The DNA eluting reagent of claim 1, wherein the combined amount of buffer, base, and chelating agent is no greater than about 20 mM, based on the total volume of the DNA eluting reagent.
11. A method for isolating DNA, comprising the steps of:
(a) contacting a biological material that contains DNA with a solid support;
(b) treating the biological material that contains DNA with a DNA purifying reagent;
(c) purifying the DNA from the remainder of the biological materials; and (d) eluting the DNA from the solid support with a DNA eluting reagent, wherein the DNA eluting reagent comprises:
(i) a buffer;
(ii) a base;
(iii) a chelating agent; and (iv) water.
12. The method of claim 11, wherein the DNA eluting reagent has a pH of at least about 7.
13. The method of claim 11, wherein the solid support is selected from a group consisting of cellulose, cellulose acetate, glass fiber, nitrocellulose, nylon, polyester, polyethersulfone, polyolefin, polyvinylidene fluoride, and combinations thereof.
14. The method of claim 13, wherein the solid support is contained in a vessel, wherein the vessel is selected from a group consisting of centrifuge tubes, spin tubes, syringes, cartridges, chambers, multiple-well plates, test tubes, and combinations thereof.
15. The method of claim 12, wherein the biological material is selected from the group consisting of eukaryotic cells, prokaryotic cells, microbial cells, bacterial cells, plant cells, mycoplasma, protoza, bacteria, fungi, viruses, and lysates and homogenates thereof.
16. The method of claim 12, wherein the biological material is selected from the group consisting of body fluids, body waste products, excretions, and tissues.
17. The method of claim 12, wherein the biological material is selected from the group consisting of environmental samples taken from air, water, sediment and soil.
18. The method of claim 11, wherein the base is chosen from the group consisting of alkaline metal hydroxides.
19. The method of claim 11, wherein the buffer is present in an amount no greater than 20 mM, based on the total volume of the DNA eluting reagent.
20. The method of claim 11, wherein the buffer is present in an amount from 0.001-20 mM, based on the total volume of the DNA eluting reagent.
21. The method of claim 11, wherein the buffer is present in an amount from 0.01-15 mM, based on the total volume of the DNA eluting reagent.
22. The method of claim 11, wherein the base is present in an amount no greater than 20 mM, based on the total volume of the DNA eluting reagent.
23. The method of claim 11, wherein the base is present in an amount from 1-10 mM, based on the total volume of the DNA eluting reagent.
24. The method of claim 11, wherein the chelating agent is present in an amount no greater than 0.1 mM, based on the total volume of the DNA eluting reagent.
25. The method of claim 11, wherein the combined amount of buffer, base, and chelating agent is no greater than about 20 mM, based on the total volume of the DNA
eluting reagent.
26. The method of claim 11, wherein the buffer is Tris, the chelating agent is EDTA, and the base is chosen from the group consisting of alkaline metal hydroxides.
27. The method of claim 11, further comprising the step of heating at greater than 60°C.
28. A kit for isolating DNA comprising:
(a) instruction means for isolating substantially pure DNA from a biological sample;
(b) a DNA purifying reagent; and (c) a DNA eluting reagent, wherein the DNA eluting reagent comprises:
(i) a buffer;
(ii) a base;
(iii) a chelating agent; and (iv) water.
29. The kit of claim 28, further comprising a solid support.
30. A kit for isolating DNA comprising:
(a) instruction means for isolating substantially pure DNA from a biological sample; and (b) a DNA eluting reagent, wherein the DNA eluting reagent comprises:
(i) a buffer;

(ii) a base;
(iii) a chelating agent; and (iv) water.
31. The kit of claim 30, further comprising a solid support.
32. A kit for isolating DNA comprising:
(a) instruction means for isolating substantially pure DNA from a biological sample;
(b) a DNA purifying reagent; and (c) a solid support treated with a lysing reagent, such that the lysing reagent is bound to the solid support.
33. The kit of claims 29, 31, and 32, further comprising a vessel for holding the solid support.
34. The kit of claim 29 and 31, wherein the solid support is treated with a lysing reagent.
35. The kit of claims 29, 31, and 32, wherein the solid support is treated with an RNA
digesting enzyme.
36. The kit of claims 29, 31, and 32, further comprising an RBC Lysis Reagent comprising ammonium chloride, sodium bicarbonate, and EDTA, for use in separating white cells from red blood cells in mammalian whole blood prior to DNA
isolation.
37. The kit of claims 29, 31, and 32, further comprising:
(a) a cell suspension reagent comprising Tris, EDTA, and sorbitol; and (b) a lytic enzyme reagent comprising a lytic enzyme, glycerol, Tris, and calcium chloride, wherein the cell suspension reagent and the lytic enzyme reagent are suitable for use in combination to digest cell walls of yeast and Gram-positive bacteria
38. The kit of claims 29, 31, and 32, further comprising reagents selected from the group consisting of:
(a) an RBC lysis reagent;
(b) a cell suspension reagent;
(c) a lytic enzyme reagent;

(d) a protein digesting reagent; and (e) an isotonic solution.
CA2319691A 1998-02-02 1999-02-02 Eluting reagents, methods and kits for isolating dna Expired - Lifetime CA2319691C (en)

Applications Claiming Priority (5)

Application Number Priority Date Filing Date Title
US12642998P 1998-02-02 1998-02-02
US12641398P 1998-02-02 1998-02-02
US60/126,429 1998-02-02
US60/126,413 1998-02-02
PCT/US1999/002190 WO1999039010A1 (en) 1998-02-02 1999-02-02 Eluting reagents, methods and kits for isolating dna

Publications (2)

Publication Number Publication Date
CA2319691A1 true CA2319691A1 (en) 1999-08-05
CA2319691C CA2319691C (en) 2012-12-18

Family

ID=26824623

Family Applications (1)

Application Number Title Priority Date Filing Date
CA2319691A Expired - Lifetime CA2319691C (en) 1998-02-02 1999-02-02 Eluting reagents, methods and kits for isolating dna

Country Status (5)

Country Link
EP (1) EP1053353A1 (en)
JP (2) JP2004500002A (en)
AU (1) AU2574299A (en)
CA (1) CA2319691C (en)
WO (1) WO1999039010A1 (en)

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AU2599800A (en) * 1999-01-06 2000-07-24 Invitrogen Corporation Methods and compositions for isolation of nucleic acid molecules
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US6803200B2 (en) 2000-12-12 2004-10-12 Invitrogen Corporation Compositions and methods for the release of nucleic acid molecules from solid matrices
US20050032105A1 (en) * 2001-10-12 2005-02-10 Bair Robert Jackson Compositions and methods for using a solid support to purify DNA
WO2003057910A2 (en) 2002-01-08 2003-07-17 Roche Diagnostics Gmbh Use of silica material in an amplification reaction
CA2501056C (en) * 2002-10-04 2012-12-11 Whatman, Inc. Methods and materials for using chemical compounds as a tool for nucleic acid storage on media of nucleic acid purification systems
JP2008220380A (en) * 2003-10-31 2008-09-25 Fujifilm Corp Method of isolating and purifying nucleic acid
US20060105372A1 (en) * 2004-11-05 2006-05-18 Bair Robert J Compositions and methods for purifying nucleic acids from stabilization reagents
US9243289B2 (en) * 2013-12-23 2016-01-26 Roche Molecular Systems, Inc. Method for screening reagents used in PCR assays
JP2017534289A (en) * 2014-11-07 2017-11-24 ザ・ジョンズ・ホプキンス・ユニバーシティ Chaotrope and volatile-free method for purifying nucleic acids from plasma
AU2016262545B2 (en) * 2015-05-12 2022-06-16 Gen-Probe Incorporated Method of pooling blood samples
MX2019010800A (en) 2017-03-15 2020-07-28 Ancestry Com Dna Llc Sample collection device and method.

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Also Published As

Publication number Publication date
JP2012157366A (en) 2012-08-23
CA2319691C (en) 2012-12-18
JP2004500002A (en) 2004-01-08
WO1999039010A1 (en) 1999-08-05
EP1053353A1 (en) 2000-11-22
AU2574299A (en) 1999-08-16

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