Room temperature elution of nucleic acids
The present invention provides compositions and methods for efficient recovery of nucleic acids, in particular DNA, from storage media. The present invention provides a method for using an alkaline elution buffer and mechanical agitation to elute nucleic acids from dried samples on storage media at room temperature. In particular, the present invention provides a method for room temperature recovery of DNA, including double stranded DNA (dsDNA) and single stranded (ssDNA), from storage media. The eluted DNA is suitable for numerous downstream applications
1 . A method for eluting nucleic acid from a nucleic acid-containing sample on storage media comprising:
(a) providing a nucleic acid-containing sample on storage media;
(b) contacting the sample on storage media with an elution buffer having a pH of between about 11.0 and about 12.0; and
(c) eluting nucleic acid from the sample on storage media.
2 . The method of claim 1 , wherein the elution buffer has a pH of between about 11.4 and about 11.8.
3 . The method of claim 1 , wherein the elution buffer comprises an organic buffering agent.
4 . The method of claim 1 , wherein the elution buffer comprises an inorganic buffering agent.
5 . The method of claim 1 , wherein the elution buffer comprises Tris (hydroxymethyl) aminomethane hydrochloride (Tris).
6 . The method of claim 5 , wherein the elution buffer comprises 10 mM Tris at a pH of about 11.4.
7 . The method of claim 5 , wherein the elution buffer comprises 10 mM Tris at a pH of about 11.5.
8 . The method of claim 5 , wherein the elution buffer comprises 10 mM Tris at a pH of about 11.6.
9 . The method of claim 5 , wherein the elution buffer comprises 10 mM Tris at a pH of about 11.7.
10 . The method of claim 5 , wherein the elution buffer comprises 10 mM Tris at a pH of about 11.8.
11 . The method of claim 1 , wherein the elution buffer comprises 4-(cyclohexylamino)-1-butanesulfonic acid (CABS).
12 . The method of claim 1 , wherein the elution buffer comprises 3-(cyclohexylamino)-1-propanesulfonic acid (CAPS).
13 . The method of claim 1 , wherein the elution buffer comprises 3-(cyclohexylamino-2-hydroxy-1-propanesulfonic acid (CAPSO).
14 . The method of claim 1 , wherein the elution buffer comprises 2-(cyclohexylamino) ethanesulfonic acid (CHES).
15 . The method of claim 1 , wherein the elution buffer comprises N-(2-hydroxyethyl)piperazine-N′-(3-propanesulfonic acid) (EPPS).
16 . The method of claim 1 , wherein the elution buffer comprises N-(2-hydroxyethyl)piperazine-N′-(2-ethanesulfonic acid (HEPES).
17 . The method of claim 1 , wherein the elution buffer comprises 2-(N-morpholino) ethanesulfonic acid (MES).
18 . The method of claim 1 , wherein the elution buffer comprises 3-(N-morpholino) propanesulfonic acid (MOPS).
19 . The method of claim 1 , wherein the elution buffer comprises piperazine-N,N′-bis (2-ethanesulfonic acid (PIPES).
20 . The method of claim 1 , wherein the elution buffer comprises [(2-hydroxy-1,1-bis[bydroxymethyl]ethyl) amino]-1-propanesulfonic acid (TAPS).
21 . The method of claim 1 , wherein the elution buffer comprises ethanolamine.
22 . The method of claim 1 , wherein the elution buffer comprises 3-amino-1-propanesulfonic acid.
23 . The method of claim 1 , further comprising contacting the sample on storage media prior with wash buffer prior to contacting the sample with elution buffer.
24 . The method of claim 23 , wherein the wash buffer comprises a buffering agent and a chelating agent.
25 . The method of claim 24 , wherein the wash buffer further comprises a detergent.
26 . The method of claim 24 , wherein the wash buffer comprises 10 mM Tris and 0.1 mM EDTA at a pH of about 8.0.
27 . The method of claim 26 , wherein the wash buffer further comprises a detergent.
28 . The method of claim 27 , wherein the wash buffer comprises 10 mM Tris, 0.1 mM EDTA, and 1% Triton X-100 at a pH of about 8.0.
29 . The method of claim 23 , wherein the wash buffer comprises an enzyme.
30 . The method of claim 29 , wherein the wash buffer comprises a protease.
31 . The method of claim 30 , wherein the wash buffer comprises proteinase K.
32 . The method of claim 23 , wherein the wash buffer comprises a lysis buffer.
33 . The method of claim 23 , wherein the wash buffer comprises a digestion buffer.
34 . The method of claim 1 , comprising eluting DNA.
35 . The method of claim 1 , comprising eluting RNA.
36 . The method of claim 1 , further comprising recovering the eluted nucleic acid.
37 . The method of claim 1 , wherein the storage media comprises cellulosic material.
38 . The method of claim 1 , wherein the storage media comprises polyester material.
39 . The method of claim 1 , wherein the storage media further comprises a detergent.
40 . The method of claim 1 , wherein the nucleic acid-containing sample is formalin-fixed paraffin-embedded tissue.
41 . A kit for eluting nucleic acid from a nucleic acid-containing sample on storage media comprising
(a) an elution buffer having a pH of between about 11.0 and about 12.0; and
(b) instruction means for eluting nucleic acid from a sample on storage media.
42 . The kit of claim 41 , wherein the elution buffer has a pH of between about 11.4 and about 11.8.
43 . The kit of claim 41 , wherein the elution buffer comprises an organic buffering agent.
44 . The kit of claim 41 , wherein the elution buffer comprises an inorganic buffering agent.
45 . The kit of claim 41 , wherein the elution buffer comprises Tris (hydroxymethyl) aminomethane hydrochloride (Tris).
46 . The kit of claim 45 , wherein the elution buffer comprises 10 mM Tris at a pH of about 11.4.
47 . The kit of claim 45 , wherein the elution buffer comprises 10 mM Tris at a pH of about 11.5.
48 . The kit of claim 45 , wherein the elution buffer comprises 10 mM Tris at a pH of about 11.6.
49 . The kit of claim 45 , wherein the elution buffer comprises 10 mM Tris at a pH of about 11.7.
50 . The kit of claim 45 , wherein the elution buffer comprises 10 mM Tris at a pH of about 11.8.
51 . The kit of claim 41 , wherein the elution buffer comprises 4-(cyclohexylamino)-1-butanesulfonic acid (CABS).
52 . The kit of claim 41 , wherein the elution buffer comprises 3-(cyclohexylamino)-1-propanesulfonic acid (CAPS).
53 . The kit of claim 41 , wherein the elution buffer comprises 3-(cyclohexylamino-2-hydroxy-1-propanesulfonic acid (CAPSO).
54 . The kit of claim 41 , wherein the elution buffer comprises 2-(cyclohexylamino) ethanesulfonic acid (CHES).
55 . The kit of claim 41 , wherein the elution buffer comprises N-(2-hydroxyethyl)piperazine-N′-(3-propanesulfonic acid) (EPPS).
56 . The kit of claim 41 , wherein the elution buffer comprises N-(2-hydroxyethyl)piperazine-N′-(2-ethanesulfonic acid (HEPES).
57 . The kit of claim 41 , wherein the elution buffer comprises 2-(N-morpholino) ethanesulfonic acid (MES).
58 . The kit of claim 41 , wherein the elution buffer comprises 3-(N-morpholino) propanesulfonic acid (MOPS).
59 . The kit of claim 41 , wherein the elution buffer comprises piperazine-N,N′-bis (2-ethanesulfonic acid (PIPES).
60 . The kit of claim 41 , wherein the elution buffer comprises [(2-hydroxy-1,1-bis[bydroxymethyl]ethyl) amino]-1-propanesulfonic acid (TAPS).
61 . The kit of claim 41 , wherein the elution buffer comprises ethanolamine.
62 . The kit of claim 41 , wherein the elution buffer comprises 3-amino-1-propanesulfonic acid.
63 . The kit of claim 41 , further comprising contacting the sample on storage media prior with wash buffer prior to contacting the sample with elution buffer.
64 . The kit of claim 63 , wherein the wash buffer comprises a buffering agent and a chelating agent.
65 . The kit of claim 64 , wherein the wash buffer further comprises a detergent.
66 . The kit of claim 64 , wherein the wash buffer comprises 10 mM Tris and 0.1 mM EDTA.
67 . The kit of claim 66 , wherein the wash buffer further comprises a detergent.
68 . The kit of claim 67 , wherein the detergent is Tween 20.
69 . The kit of claim 67 , wherein the detergent is Triton X-100.
70 . The kit of claim 69 , wherein the wash buffer comprises 10 mM Tris, 0.1 mM EDTA, and 1% Triton X-100.
71 . The kit of claim 63 , wherein the wash buffer comprises a protease.
72 . The kit of claim 63 , wherein the wash buffer comprises a lysis buffer.
73 . The kit of claim 63 , wherein the wash buffer comprises a digestion buffer.
74 . The kit of claim 41 , further comprising means for recovering the eluted nucleic acid.
75 . The kit of claim 74 , comprising means for recovering DNA.
76 . The kit of claim 74 , comprising means for recovering RNA.
75 . The kit of claim 74 , further comprising means for analyzing the eluted nucleic acid.
76 . A method for eluting nucleic acid from a human blood sample on storage media comprising:
(a) providing a human blood sample on storage media;
(b) contacting the blood sample on storage media with a wash buffer;
(c) removing the wash buffer;
(d) contacting the sample on storage media with an elution buffer comprising 10 mM Tris having a pH of between about 11.5 and about 11.7;
(e) eluting nucleic acid from the blood sample on storage media; and
(f) recovering eluted nucleic acid.
77 . The method of claim 76 , wherein the wash buffer comprises 10 mM Tris, 0.1 mM EDTA, and 1% Triton X-100 at a pH of about 8.0.
78 . The method of claim 76 , wherein the wash buffer comprises proteinase K.
79 . The method of claim 76 , comprising eluting DNA from a blood sample on storage media.