Isolation and purification of nucleic acids
AssigneeCortex Biochem, Inc.
InventorsRuhangiz D. Nargessi, Matt Pourfarzaneh
Life of this patent
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Abstract
A method for the isolation and purification of nucleic acids such as DNA, RNA, and PNA from various sources using cellulose particles or cellulose paper. Adjusting the concentrations of the salt and polyalkylene glycol to the levels that result in binding of nucleic acids to the cellulose particles or cellulose paper. Separating the nucleic acids bound to the cellulose particles or paper and eluting the nucleic acids from the particles or paper.
Claims33
A method to bind nucleic acids to cellulose, wherein said nucleic acids have not been purified or semi-purified, said method comprising:
- a) combining cellulose, wherein said cellulose is not chemically coupled or physically adsorbed to another material, with a solution containing said nucleic acids, thereby producing a combination, and
- b) adjusting the salt and polyalkylene glycol concentrations of the combination to final concentrations suitable for binding the nucleic acids to the cellulose using a binding buffer having a concentration of from 0.5 M to 1.5 M salt and from 5% to 15% polyalkylene glycol, whereby all or a portion of the nucleic acids in the solution binds to the cellulose, and wherein the nucleic acids bind to the cellulose while the nucleic acids are in a non-aggregated state.
A method of separating nucleic acids from non-nucleic acid materials in a nucleic acid solution, wherein said nucleic acids have not been purified, said method comprising:
- a) combining cellulose particles with a solution containing said nucleic acids and non-nucleic acid materials to produce a first combination;
- b) adjusting the salt and polyethylene glycol concentrations of the first combination to concentrations suitable for binding said nucleic acids in the solution to the cellulose particles using a binding buffer having a concentration of from 0.5 M to 1.5 M salt and from 5% to 15% polyethylene glycol, producing a second combination comprising cellulose-bound nucleic acids, and wherein the nucleic acids bind to the cellulose while the nucleic acids are in a non-aggregated state;
- c) separating the cellulose-bound nucleic acids from the second combination;
- d) contacting the cellulose-bound nucleic acids separated in c) with an elution buffer to release the bound nucleic acids from the cellulose particles and into the elution buffer;
- and
- e) separating the cellulose particles from the elution buffer to provide nucleic acids that are substantially free of the non-nucleic acid materials, wherein the separation of the cellulose particles in step c) and e) is carried out using a centrifuge or a column.
A method to bind nucleic acids to cellulose paper, wherein said nucleic acids have not been purified or semi-purified, said method comprising:
- a) combining cellulose paper with a solution containing said nucleic acids, thereby producing a combination, and
- b) adjusting the salt and polyalkylene glycol concentrations of the combination to concentrations suitable for binding the nucleic acids to the cellulose paper using a binding buffer having a concentration of from 0.5 M to 1.5 M salt and from 5% to 15% polyalkylene glycol, whereby all or a portion of the nucleic acids in the solution binds to the cellulose paper, and wherein the nucleic acids bind to the cellulose while the nucleic acids are in a non-aggregated state.
A method of separating nucleic acids from non-nucleic acid materials, wherein said nucleic acids have not been purified or semi-purified, said method comprising:
- a) combining cellulose paper with a solution containing said nucleic acids and non-nucleic acid materials to provide a first combination;
- b) adjusting the salt and polyethylene glycol concentrations of the first combination to concentrations suitable for binding said nucleic acids to the cellulose paper using a binding buffer having a concentration of from 0.5 M to 1.5 M salt and from 5% to 15% polyethylene glycol, producing a second combination comprising cellulose paper-bound nucleic acids, and wherein the nucleic acids bind to the cellulose while the nucleic acids are in a non-aggregated state;
- c) separating the cellulose paper-bound nucleic acids from the second combination;
- d) contacting the cellulose paper-bound nucleic acids separated in c) with an elution buffer to release the bound nucleic acids from the cellulose paper and into the elution buffer;
- and
- e) separating the cellulose paper from the elution buffer to provide nucleic acids that are substantially free of the non-nucleic acid materials.
A method for binding nucleic acids to cellulose, comprising combining cellulose, wherein said cellulose is not chemically coupled or physically adsorbed to another material, with nucleic acids that have not been purified or semi-purified in a mixture containing sodium chloride at a concentration of from about 0.7 M to 2.5 M and a polyethylene glycol at a concentration of from about 5% to 10%, said concentrations being sufficient to bind said nucleic acids to said cellulose while the nucleic acids are in a non-aggregated state.
Dependencies are inferred from the claim text.