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1. WO1995006652 - COMPOSITIONS ET PROCEDE DE PURIFICATION D'ACIDES NUCLEIQUES

Note: Texte fondé sur des processus automatiques de reconnaissance optique de caractères. Seule la version PDF a une valeur juridique

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CLAIMS
1. A mixture of silica gel and glass particles.

2. A mixture of Claim 1 which consists
essentially of silica gel and glass microfibers.

3. A mixture of Claim 1 wherein the ratio by weight of silica gel to glass particles is 5:1 to 50:1.

4. A mixture of Claim 2 wherein the ratio by weight of silica gel to glass microfibers is 5:1 to 50:1.

5. A mixture of Claim 1 further mixed with a chaotropic salt.

6. A mixture of Claim 2 further mixed with a chaotropic salt.

7. A mixture of Claim 3 further mixed with a chaotropic salt.

8. A mixture of Claim 4 further mixed with a chaotropic salt.

9. A mixture of any one of Claims 5 - 8 wherein the chaotropic salt is guanidinium chloride.

10. A mixture of any one of Claims 5 - 8 wherein the chaotropic salt is guanidinium thiocyanate.

11. A mixture of Claim 10 further mixed with a chelating agent.

12. A mixture of Claim 11 wherein the chelating agent is selected from the group consisting of EGTA and a soluble salt of EDTA.

13. A suspension of silica gel and glass
particles in an aqueous solution, which comprises one or more chaotropic salts and has a concentration of
chaotropic ions of at least 4 M.

14. A suspension of Claim 13 wherein the glass particles are glass microfibers.

15. A suspension of Claim 13 wherein the weight ratio of silica gel to glass particles is 5:1 to 50:1.

16. A suspension of Claim 14 wherein the weight ratio of silica gel to glass particles is 5:1 to 50:1.

17. A suspension of Claim 13 wherein, in the solution, guanidinium ion is present at a concentration of at least 4 M.

18. A suspension of Claim 14 wherein, in the solution, guanidinium ion is present at a concentration of at least 4 M.

19. A suspension of Claim 15 wherein, in the solution, guanidinium ion is present at a concentration of at least 4 M.

20. A suspension of Claim 16 wherein, in the solution, guanidinium ion is present at a concentration of at least 4 M.

21. A suspension of Claim 17 wherein the chaotropic salt is guanidinium hydrochloride at a concentration of 6 M to 8 M.

22. A suspension of Claim 18 wherein the chaotropic salt is guanidinium hydrochloride at a concentration of 6 M to 8 M.

23. A suspension of Claim 19 wherein the chaotropic salt is guanidinium hydrochloride at a concentration of 6 M to 8 M.

24. A suspension of Claim 20 wherein the chaotropic salt is guanidinium hydrochloride at a concentration of 6 M to 8 M.

25. A suspension of Claim 17 wherein the chaotropic salt is guanidinium thiocyanate at a concentration of 5 M to 7 M.

26. A suspension of Claim 18 wherein the chaotropic salt is guanidinium thiocyanate at a concentration of 5 M to 7 M.

27. A suspension of Claim 19 wherein the chaotropic salt is guanidinium thiocyanate at a concentration of 5 M to 7 M.

28. A suspension of Claim 20 wherein the chaotropic salt is guanidinium thiocyanate at a concentration of 5 M to 7 M.

29. A suspension according to any of
Claims 24 - 28 which comprises EGTA or a salt of EDTA.

30. A suspension according to Claim 29 which comprises the disodium salt of EDTA at a concentration between about 5 mM and about 15 mM.

31. A method of separating DNA or RNA, which has a length of greater than 50 bases, from a first aqueous solution comprising DNA or RNA, resoectively, of such length, which method comprises (1) preparing a second aqueous solution by mixing an aliquot of said first aqueous solution with a third aqueous solution, which comprises one or more chaotropic salts and has a concentration of chaotropic ions of at least 4 M, provided that the volume of said third aqueous solution is less than about twice the volume of said aliquot of said first aqueous solution; and (2) contacting an aliquot of said second aqueous solution with a mixture of silica gel and glass particles.

32. A method of Claim 31 which comprises, after step (2) , (3) separating from a first portion of the mixture of silica gel and glass particles, which was contacted in step (2) with said aliquot of second solution, substantially all of said aliquot which is not bound to said first portion of said mixture;
(4) washing a second portion of said mixture of silica gel and glass particles, which is a portion of said first portion of said mixture, with a fourth aqueous solution which is effective to remove substantially all of the chaotropic ions, but substantially ineffective in removing DNA other than short oligonucleotides, from said second portion of said mixture of silica gel and glass particles; and, after step (4) , (5) washing a third portion of said mixture of silica gel and glass particles, which is a portion of said second portion of said mixture, with water or a fifth aqueous solution to elute DNA from said third portion.

33. A method according to Claim 31 wherein, in the third solution, the chaotropic salts are selected from the group consisting of guanidinium hydrochloride and guanidinium thiocyanate; the mixture of silica gel and glass particles consists essentially of silica gel and glass microfibers in a ratio by weight of 5 parts to 50 parts silica gel to 1 part glass microfibers.

34. A method according to Claim 32 wherein, in the third solution, the chaotropic salts are selected from the group consisting of guanidinium hydrochloride and guanidinium thiocyanate; the mixture of silica gel and glass particles consists essentially of silica gel and glass microfibers in a ratio by weight of 5 parts to 50 parts silica gel to 1 part glass microfibers; the fourth solution is an aqueous solution comprising 20 % by volume to 95 % by volume of an alcohol selected from the group consisting of methanol, ethanol and
isopropanol; and the fifth solution is a low salt buffer with a pH between 6.5 and 8.5.

35. A method according to Claim 33 wherein, in the third solution, the chaotropic salt is guanidinium hydrochloride at a concentration between 6 M and 8 M or guanidinium thiocyanate at a concentration between 5 M and 7 M and, if the chaotropic salt is guanidinium thiocyanate, the third solution comprises EDTA at a concentration between 5 mM and 15 mM.

36. A method according to Claim 34 wherein, in the third solution, the chaotropic salt is guanidinium hydrochloride at a concentration between 6 M and 8 M or guanidinium thiocyanate at a concentration between 5 M and 7 M and, if the chaotropic salt is guanidinium thiocyanate, the third solution comprises EDTA at a concentration between 5 mM and 15 mM.

37. A method according to Claim 36 wherein the fifth solution is TE buffer.

38. A method according to Claim 31 or 32 wherein the step of preparing the second solution and the step of contacting the second solution with the mixture of silica gel and glass particles are carried out
simultaneously by combining an aliquot of the first solution with a suspension of silica gel and glass particles in an aqueous solution, which comprises one or more chaotropic salts and has a concentration of
chaotropic ions of at least 4 M, provided that the volume of solution in said suspension is less than about twice the volume of said aliquot of said first solution.

39. A method according to Claim 33 or 34 wherein the step of preparing the second solution and the step of contacting the second solution with the mixture of silica gel and glass particles are carried out
simultaneously by combining an aliquot of the first solution with a suspension of silica gel and glass particles in an aqueous solution, which comprises one or more chaotropic salts and has a concentration of
chaotropic ions of at least 4 M, provided that the volume of solution in said suspension is less than about twice the volume of said aliquot of said first solution.

40. A method according to Claim 35, 36 or 37 wherein the step of preparing the second solution and the step of contacting the second solution with the mixture of silica gel and glass microfibers are carried out simultaneously by combining an aliquot of the first solution with an aliquot of a suspension of diatomaceous earth and glass microfibers in an aqueous solution, which comprises guanidinium hydrochloride at a
concentration between 6 M and 8 M or guanidinium
thiocyanate at a concentration between 5 M and 7 M, provided that the volume of solution in said aliquot of said suspension is less than about twice the volume of said aliquot of said first solution.

41. A method according to any one of
Claims 31 - 40 wherein the separated nucleic acid is DNA.

42. A method according to any one of
Claims 31 - 40 wherein the separated nucleic acid is RNA.