Method for isolating and purifying nucleic acids
AssigneeHitachi, Ltd.
InventorsKyoko Kojima, Satoshi Ozawa
Life of this patent
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Abstract
A method for isolating and purifying nucleic acids with an improved recovery yield is provided. A mixed solution containing the nucleic acids, salts, and an organic solvent is contacted with an adsorption support to cause the nucleic acids to be adsorbed on the support. Then, the nucleic acids are desorbed from the support using an elution bugger. At least one compound containing 2 to 10 carbon atoms as selected from the group consisting of aliphatic ether, aliphatic ester, and aliphatic ketone is used as the organic solvent. The method improves the yield of nucleic acids collection, is easy to implement and less susceptible to contamination.
Claims20
A method for extracting nucleic acids comprising:
- setting a first mixed liquid at a certain temperature, wherein the first mixed liquid containing a protease, a chaotropic salt, a surfactant and a sample;
- adding Diethylene glycol dimethyl ether to the first mixed liquid to get a second mixed liquid after the step of setting a first mixed liquid;
- absorbing nucleic acids, contained in the second mixed liquid, to a support:
- washing the support with a solution containing ethanol;
- recovering the nucleic acids absorbed to the support, by eluting from the support.
A reagents kit for use in extracting nucleic acids, comprises
- a protease,
- a first liquid containing a chaotropic salt and a surfactant;
- a second liquid containing Diethylene glycol dimethyl ether;
- a support for absorbing nucleic acids;
- a third liquid containing ethanol;
- an elution buffer for eluting the nucleic acids, absorbed to the support, from the support.
A method for extracting nucleic acids from blood comprising:
- setting a first mixed liquid at a certain temperature, wherein the first mixed liquid containing a protease, a chaotropic salt, a surfactant and a sample,
- adding at least one of compounds which are any of aliphatic ether, aliphatic ester, and aliphatic ketone, containing 2 to 10 carbon atoms to the first mixed liquid to get a second mixed liquid after the step of setting a first mixed liquid;
- absorbing nucleic acids, contained in the second mixed liquid, to a support:
- washing the support with a solution containing ethanol;
- recovering the nucleic acids, absorbed to the support, by eluting from the support.
A reagents kit for use in extracting nucleic acids from blood, comprises
- a protease,
- a first liquid containing a chaotropic salt and a surfactant;
- a second liquid containing at least one of compounds which are any of aliphatic ether, aliphatic ester, and aliphatic ketone, containing 2 to 10 carbon atoms;
- a support for absorbing nucleic acids,
- a third liquid containing ethanol;
- an elution buffer for eluting the nucleic acids, absorbed to the support, from the support.
A method for extracting nucleic acids comprising:
- lysing cells for getting a first mixed liquid;
- adding at least one of compounds which are any of aliphatic ether, aliphatic ester, and aliphatic ketone, containing 2 to 10 carbon atoms to the first mixed liquid for getting a second mixed liquid;
- absorbing nucleic acids, contained in the second mixed liquid, to a support by letting the second mixed liquid passing through a vessel containing the support both a way by sucking force and a substantially opposite way;
- washing the support;
- eluting the nucleic acids, absorbed to the support, from the support.
A method for extracting nucleic acids comprising:
- lysing cells for getting a first mixed liquid;
- adding at least one of compounds which are any of aliphatic ether, aliphatic ester, and aliphatic ketone, containing 2 to 10 carbon atoms to the first mixed liquid for getting a second mixed liquid;
- transferring the second mixed liquid into a column containing a support;
- absorbing nucleic acids, contained in the second mixed liquid, to a support by sucking the second mixed liquid from the column;
- washing the support by injecting a washing solution into the column and sucking the washing solution from the column;
- eluting the nucleic acids, absorbed to the support, from the support by injecting a eluting solution into the column and sucking the eluting solution from the column.
A reagents kit for use in extracting nucleic acids, comprises
- a proteolytic enzyme,
- a reaction solution containing a chaotropic salt and a surfactant and supplied into a sample;
- an additive solution containing at least one of compounds which are any of aliphatic ether, aliphatic ester, and aliphatic ketone, containing 2 to 10 carbon atoms, added into the reaction solution supplied into the sample;
- a support for absorbing nucleic acids;
- a washing buffer for washing the support;
- an elution buffer for eluting the nucleic acids, absorbed to the support, from the support.
A reagents kit for use in extracting nucleic acids, comprises
- a proteinase K,
- a first liquid containing Guanidine hydrochloride and Polyoxyethylne(10) isooctylphenyl ether;
- a second liquid containing Diethylene glycol dimethyl ether;
- a support for absorbing nucleic acids;
- a third liquid containing potassium acetate and ethanol;
- an elution solution for eluting the nucleic acids, absorbed to the support, from the support.
Dependencies are inferred from the claim text.