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US 2010/0160619 A1Published application
Compositions and methods for using a solid support to purify dna
AssigneeQiagen North American Holdings, Inc.
InventorsRobert Jackson Bair, Kristen Campbell Benedict, Wendy J. Kivens, Robert W. Kwiatkowski, JR., Kim Paulsen, Daniel A. Strom, John M. Wages
Life of this patent
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Abstract
Reagents, methods and kits for the purification of DNA from biological materials are provided.
Claims32
-18. (canceled)
A method for isolating substantially pure and undegraded DNA from biological material, comprising the steps of:
- (a) contacting the biological material with a DNA Lysing Solution to form a mixture, wherein the DNA Lysing Solution is buffered at pH of greater than 7, and wherein the DNA Lysing Solution comprises a lithium salt and at least one surfactant;
- (b) contacting the mixture with a DNA Spiking Solution;
- (c) contacting the mixture to a solid support such that DNA present in the biological material binds to the solid support;
- (d) washing the solid support with a Wash Solution;
- and
- (e) eluting the DNA with a DNA Elution Solution.
3 more elements
A method for purifying substantially pure and undegraded DNA from biological material, comprising the steps of:
- (a) contacting a biological material containing DNA with a solid support pre-treated with a DNA Lysing Solution, wherein the DNA Lysing Solution is buffered at a pH of greater than about 7, and wherein the DNA Lysing Solution comprises a surfactant and a lithium salt;
- (b) adding a DNA Spiking Solution to the biological material;
- (c) contacting the biological material to the solid support in order to release nucleic acids comprising substantially undegraded DNA and non-nucleic acid biological matter, wherein the nucleic acids comprising substantially undegraded DNA bind to the solid support;
- (d) washing the solid support with a Wash Solution to remove biological materials other than bound nucleic acids comprising undegraded DNA;
- and
- (e) eluting the bound undegraded DNA from the solid support with a DNA Elution Solution.
3 more elements
A direct lysis method for purifying substantially pure and undegraded DNA from biological material without using a red blood cell lysis step, comprising:
- (a) contacting a biological material containing DNA, with a first DNA Lysing Solution, wherein the first DNA Lysing Solution is buffered at a pH of greater than about 7 and comprises a surfactant at a concentration of between about 5-15% v/v and a DNA-complexing salt at a concentration of greater than 1 M,
- (b) contracting the biological material with a second DNA Lysing Solution, wherein the second DNA Lysing Solution comprises a surfactant and a DNA-complexing salt greater than 1 M;
- (c) contacting the mixture with a DNA Spiking Solution;
- (d) contacting the mixture with a solid support, wherein nucleic acids comprising substantially undegraded DNA from the biological material bind to the solid support;
- (e) washing the solid support with a DNA Wash Solution to remove biological materials other than bound nucleic acids comprising substantially undegraded DNA, the DNA Wash Solution comprising a DNA-complexing salt and an alcohol;
- and
- (f) preferentially eluting the bound substantially undegraded DNA from the solid support with a DNA Elution Solution.
4 more elements
A method for purifying substantially pure and undegraded DNA from biological material comprising:
- (a) contacting a biological material containing DNA with DNA Lysing Solution buffered at a pH of greater than about 7, wherein the DNA Lysing Solution comprises a surfactant and DNA-complexing salt at a concentration greater than 1 M;
- (b) contacting the biological material to a solid support, wherein nucleic acids comprising substantially undegraded DNA bind to the solid support;
- (c) contacting the solid support with a DNA Spiking Solution;
- (d) washing the solid support with a DNA Wash Solution to remove biological materials other than bound nucleic acids comprising undegraded DNA, the DNA Wash Solution comprising a DNA-complexing salt and an alcohol;
- and
- (e) preferentially eluting bound substantially undegraded DNA from the solid support with a DNA Elution Solution.
3 more elements
A method for purifying substantially pure and undegraded DNA fixed cervical cell samples, comprising the steps of
- (a) contacting a biological material containing DNA with DNA Lysing Solution buffered at a pH of greater than about 7, wherein the DNA Lysing Solution comprises a surfactant at a concentration of between about 25-35% v/v, a chelating agent, and a DNA-complexing salt of greater than 1 M;
- (b) adding to the mixture, an optional DNA Spiking Solution containing alcohol;
- (c) contacting the biological material to the solid support in order to release nucleic acids comprising substantially undegraded DNA and non-nucleic acid biological matter causing nucleic acids comprising substantially undegraded DNA to bind to the solid support;
- (d) washing the solid support with a DNA Wash Solution to remove biological materials other than bound nucleic acids comprising undegraded DNA;
- and
- (e) preferentially eluting the bound undegraded DNA from the solid support with a DNA Elution Solution in order to obtain substantially pure and undegraded DNA.
3 more elements
Dependencies are inferred from the claim text.