Compositions, methods, and kits for isolating nucleic acids using surfactants and proteases
First Claim
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1. A composition for macerating whole tissue, wherein the whole tissue is not a microorganism, a virus, or blood, comprising:
- at least one cationic surfactant, at least one protease, a buffer, and a salt at a concentration of about 550 mM or less, wherein the cationic surfactant accelerates maceration of the whole tissue by the at least one protease.
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Abstract
The invention provides compositions and methods for releasing and for isolating nucleic acids from biological samples, preferably from whole tissue, using cationic surfactants and proteases. The surfactant-protease combinations, when used with whole tissue, macerate the tissue, lyse individual cells, release nucleic acids, and inactivate nucleases. Kits for isolating nucleic acids from biological samples, particularly from whole tissue, are also provided.
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47 Claims
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1. A composition for macerating whole tissue, wherein the whole tissue is not a microorganism, a virus, or blood, comprising:
- at least one cationic surfactant, at least one protease, a buffer, and a salt at a concentration of about 550 mM or less, wherein the cationic surfactant accelerates maceration of the whole tissue by the at least one protease.
- View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25)
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26. A composition for macerating whole tissue, wherein the whole tissue is not a microorganism, a virus, or blood, comprising:
- at least one cationic surfactant, at least one protease, a buffer, and the whole tissue, wherein the at least one cationic surfactant is selected from;
a) an alkyltrimethyl ammonium salt having the structure;
wherein R1, R2, and R3 are methyl groups, and R4 is an alkyl group, comprising 6, 8, 10, 12, 14, 16, or 18 carbon atoms; and b) a benzyldimethyl-n-alkylammonium salt having the structure;
wherein R1 and R2 are methyl groups. R3 is an aryl group comprising six carbon atoms, and R4 is an alkyl group comprising 6, 8, 10, 12, 14, 16, or 18 carbon atoms. - View Dependent Claims (27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47)
- at least one cationic surfactant, at least one protease, a buffer, and the whole tissue, wherein the at least one cationic surfactant is selected from;
Specification