Methods and compositions for extraction and storage of nucleic acids
View Patent ↗The present disclosure generally relates to solid matrices for the extraction, stabilization, and storage of nucleic acids, particularly RNA, in a dry format under ambient conditions for a prolonged period of time. Methods for extracting, collecting, and recovering nucleic acids from the solid compositions are also described.
1. A solid matrix for extraction and storage of nucleic acids from a biological sample, wherein the solid matrix is impregnated with a at least one protein denaturant, at least one reducing agent, a UV protectant and a buffer present in the solid matrix in a dry state and where the solid matrix is a porous matrix comprising cellulose, cellulose acetate, glass fiber, or any combination thereof; and where
the protein denaturant is guanidinium thiocyanate;
the reducing agent is tris(2-carboxyethyl)phosphine (TCEP); and
the UV protectant is THQ, MEHQ or a combination thereof.
2. The solid matrix of claim 1 , wherein the composition present in the solid matrix further comprises an RNase inhibitor.
3. The solid matrix of claim 1 , wherein the solid matrix permits prolonged storage of nucleic acids in a dry format under ambient conditions.
4. The solid matrix of claim 3 , wherein the nucleic acids are RNA, DNA, or a combination thereof.
5. The solid matrix of claim 4 , wherein the nucleic acids are RNA.
6. The solid matrix of claim 1 , wherein the porous matrix is a cellulose paper.
7. The solid matrix of claim 1 , wherein the buffer is selected from the group consisting of 2-Amino-2-hydroxymethyl-propane-1,3-diol (Tris), 2-(N-morpholino)ethanesulfonic acid (MES), 3-(N-morpholino)propanesulfonic acid (MOPS), 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), a citrate buffer, and a phosphate buffer.
8. The solid matrix of claim 7 , wherein the buffer has a pH range of between 3 and 6.
9. The solid matrix of claim 2 , wherein the RNase inhibitor is vanadyl ribonucleoside complex (VRC), or a commercially available RNase inhibitor.
10. A method for extracting and storing nucleic acids from a sample comprising:
a) providing a solid matrix, wherein a composition comprising at least one protein denaturant, at least one reducing agent, a UV protectant, and a buffer, are present in the solid matrix in a dry state and where the solid matrix is a porous matrix comprising cellulose, cellulose acetate, glass fiber, or any combination thereof where;
the protein denaturant is guanidinium thiocyanate;
the reducing agent is tris(2-carboxyethyl)phosphine (TCEP); and
the UV protectant is THQ, MEHQ or a combination thereof;
b) applying a biological sample to the solid matrix to collect the nucleic acids;
c) drying the solid matrix; and
d) storing the nucleic acids on the solid matrix in a dry state under ambient conditions.
11. The method claim 10 , wherein the method further comprises recovering the nucleic acids from the solid matrix.
12. The method of claim 10 , wherein the biological sample is blood, serum, tissue, saliva, or cells.
13. The method of claim 10 , wherein the sample is a purified nucleic acid sample or a tissue culture cell preparation.
14. The method of claim 10 , wherein purification of the nucleic acids from the sample is not required prior to applying the sample to the solid matrix for stabilizing and preserving the nucleic acids.
15. The method of claim 10 , wherein the method permits the prolonged storage of RNA in a dry format under ambient conditions.
16. The method of claim 11 , wherein the nucleic acids are recovered from the solid matrix by rehydrating the matrix in an aqueous solution, a buffer, or an organic solution, extracting the nucleic acid from the matrix, and wherein the nucleic acids are subjected to further analysis.
17. The method of claim 11 , wherein the nucleic acids are recovered from the solid matrix by electroelution.