Compositions, methods, and kits for isolating nucleic acids using surfactants and proteases
View Patent ↗The invention relates to compositions and methods for isolating nucleic acids from biological samples, including whole tissue. The invention also provides kits for isolating nucleic acids from biological samples.
1. A method for isolating nucleic acids from a biological sample comprising:
combining the sample with at least one cationic surfactant, at least one protease, and a buffer, to form a reaction composition;
incubating the reaction composition at a temperature suitable for releasing nucleic acid from the biological sample; and
isolating the released nucleic acid;
wherein the method further comprises adding a polymer.
2. The method of claim 1 , wherein the polymer is a polyethylene glycol.
3. A method for isolating nucleic acids from a biological sample comprising:
combining the sample with at least one cationic surfactant, at least one protease, and a buffer, to form a reaction composition;
incubating the reaction composition at a temperature suitable for releasing nucleic acid from the biological sample; and
isolating the released nucleic acid;
wherein the method further comprises adding a divalent cation capable of precipitating the nucleic acid, wherein the divalent cation is zinc.
4. A method for isolating nucleic acids from a biological sample comprising:
combining the samnie with at least one cationic surfactant, at least one protease, a buffer and a ribonuclease inhibitor, to form a reaction composition;
incubating the reaction composition at a temperature suitable for releasing nucleic acid from the biological sample; and
isolating the released nucleic acid;
wherein the nucleic acid is ribonucleic acid.
5. The method of claim 4 , wherein the ribonuclease inhibitor comprises aurintricarboxylic acid, vanadylate ribonucleoside complexes, phenylglyoxal, p-hydroxyphenylglyoxal, polyamines, spermidine, 9-aminoacridine, iodoacetate, bentonite, poly[2′-O-(2,4-dinitrophenyl)]poly(adenyhlic acid), zinc sulfate, bromopyruvic acid, formamide, copper, or zinc.
6. The method of claim 5 , wherein the ribonuclease inhibitor is aurintricarboxylic acid.
7. A method for isolating nucleic acids from a biological sample comprising:
combining the sample with at least one cationic surfactant, at least one protease, and a buffer, to form a reaction composition;
incubating the reaction composition at a temperature suitable for releasing nucleic acid from the biological sample, wherein the reaction composition is incubated at a temperature of less than 60° C.; and
isolating the released nucleic acid;
wherein the nucleic acid is ribonucleic acid.
8. The method of claim 7 , wherein the reaction composition is incubated at a temperature between 40° C. and 50° C.
9. A method for isolating nucleic acids from a biological sample comprising:
combining the sample with at least one cationic surfactant, at least one protease, and a buffer, to form a reaction composition;
incubating the reaction composition at a temperature suitable for releasing nucleic acid from the biological sample, wherein the reaction composition has a pH between 5.0 and 7.0; and
isolating the released nucleic acid;
wherein the nucleic acid is ribonucleic acid.
10. The method of claim 9 , wherein the reaction composition is incubated at a temperature between 40° C. and 50° C., and wherein the reaction composition further comprises aurintricarboxylic acid.
11. A method for releasing nucleic acids from a biological sample comprising:
combining the sample with at least one cationic surfactant, at least one protease, a buffer and a ribonuclease inhibitor, to form a reaction composition; and
incubating the reaction composition at a temperature suitable for releasing the nucleic acids from the biological sample;
wherein the nucleic acid is ribonucleic acid.
12. The method of claim 11 , wherein the ribonuclease inhibitor comprises aurintricarboxylic acid, vanadylate ribonucleoside complexes, phenylglyoxal, p-hydroxyphenylglyoxal, polyamines, spermidine, 9-aminoacridine, iodoacetate, bentonite, poly[2′-O-(2,4-dinitrophenyl)]poly(adenyhlic acid), zinc sulfate, bromopyruvic acid, formamide, copper, or zinc.
13. The method of claim 12 , wherein the ribonuclease inhibitor is aurintricarboxylic acid.
14. A method for releasing nucleic acids from a biological sample comprising:
combining the sample with at least one cationic surfactant, at least one protease, and a buffer, to form a reaction composition; and
incubating the reaction composition at a temperature suitable for releasing the nucleic acids from the biological sample, wherein the reaction composition is incubated at a temperature of less than 60° C.;
wherein the nucleic acid is ribonucleic acid.
15. The method of claim 14 , wherein the reaction composition is incubated at a temperature between 40° C. and 50° C.
16. A method for releasing nucleic acids from a biological sample comprising:
combining the sample with at least one cationic surfactant, at least one protease, and a buffer, to form a reaction composition, wherein the reaction composition has a pH between 5.0 and 7.0; and
incubating the reaction composition at a temperature suitable for releasing the nucleic acids from the biological sample;
wherein the nucleic acid is ribonucleic acid.
17. The method of claim 16 , wherein the reaction composition is incubated at a temperature between 40° C. and 50° C., and wherein the reaction composition further comprises aurintricarboxylic acid.
18. A kit for obtaining nucleic acid from a biological sample comprising at least one cationic surfactant; at least one protease; and at least one organic solvent for extracting the nucleic acids, precipitating the nucleic acids, or both extracting and precipitating the nucleic acids; wherein the organic solvent for extracting nucleic acids comprises phenol and the organic solvent for precipitating nucleic acids comprises isopropanol or ethanol.
19. A kit for obtaining nucleic acid from a biological sample comprising at least one cationic surfactant and at least one protease, wherein the kit further comprises a polymer.
20. A kit for obtaining nucleic acid from a biological sample comprising at least one cationic surfactant and at least one protease, wherein the kit further comprises at least one ribonuclease inhibitor.
21. The kit of claim 20 , wherein the at least one ribonuclease inhibitor is aurintricarboxylic acid, vanadylate ribonucleoside complexes, phenylglyoxal, p-hydroxyphenylglyoxal, polyamines, spermidine, 9-aminoacridine, iodoacetate, Bentonite, poly[2′-O-(2,4-dinitrophenyl)]poly(adenyhlic acid), zinc sulfate, bromopyruvic acid, formamide, copper, or zinc.
22. A kit for obtaining nucleic acid from a biological samDle comprising at least one cationic surfactant and at least one protease, wherein the kit further comprises at least one solubilizing agent.
23. The kit of claim 22 , wherein the solubilizing agent comprises 1-methyl 2 pyrolidinone, N-methyl pyrolidinone, pyrolidinone, dimethylformamide, or dimethylsulfoxide.