Archiving of vectors
View Patent ↗The invention relates to a solid medium or matrix for storage of nucleic acid molecules (e.g. RNA and/or DNA), particularly vectors and especially plasmids, comprising a solid matrix preferably having a compound or composition which protects against degradation of nucleic acids incorporated into or absorbed on the matrix. The invention also relates to methods for storage or isolation/purification of nucleic acids using this solid medium, and in situ use of the stored or isolated/purified nucleic acids.
1. A method of isolating or purifying one or more plasmids from a host cell comprising:
a) providing a dry matrix or solid medium, wherein said dry matrix or solid medium further comprises:
i) a weak base;
ii) a chelating agent; and
iii) an anionic surfactant or an anionic detergent;
b) contacting the matrix or solid medium with a sample comprising a host cell containing said plasmid or plasmids;
c) releasing said one or more plasmids from said host cell and onto said matrix or solid medium; and
d) isolating all or a portion of said plasmid or plasmids from said matrix or solid medium.
2. The method of claim 1 , wherein said matrix or solid medium protects against degradation of said plasmid or plasmids.
3. The method of claim 1 , wherein said matrix or solid medium comprises a polymeric matrix or solid medium.
4. The method of claim 3 , wherein the polymeric matrix or solid medium comprises a cellulose-based matrix.
5. The method of claim 3 , wherein the polymeric matrix or solid medium comprises a micromesh synthetic polymer matrix.
6. The method of claim 5 , wherein the polymeric matrix or solid medium comprises a micromesh synthetic plastic matrix.
7. The method of claim 1 , wherein said matrix or solid matrix is contacted with said one or more cells in solution.
8. The method of claim 1 , wherein said matrix or solid medium further comprises uric acid or a urate salt.
9. The method of claim 1 , wherein said host cells comprise eukaryotic cells.
10. The method of claim 1 , wherein said host cells comprise prokaryotic cells.
11. The method of claim 10 , wherein said prokaryotic cells comprise bacterial cells.
12. A method of isolating or purifying one or more plasmids from a host cell comprising:
a) contacting a solid medium with a sample comprising a host cell containing said plasmid, wherein the solid medium comprises:
i) a polymeric matrix comprising a cellulose-based matrix, a micromesh synthetic polymer matrix, or a micromesh synthetic plastic matrix;
ii) a weak base;
iii) a chelating agent; and
iv) an anionic surfactant or an anionic detergent;
b) releasing the plasmid from the host cell and onto said medium; and
c) isolating said plasmid from said medium.
13. The method of claim 12 , wherein said solid medium further comprises uric acid or a urate salt.
14. A method of isolating or purifying one or more plasmids from a host cell comprising:
a) contacting a solid medium with a sample comprising a host cell containing said plasmid, wherein said solid medium protects against degradation of said plasmid and wherein the solid medium comprises:
i) a polymeric matrix comprising a cellulose-based matrix, a micromesh synthetic polymer matrix, or a micromesh synthetic plastic matrix; and
ii) a composition sorbed to the polymeric matrix, wherein the composition comprises:
a weak base;
a chelating agent; and
an anionic surfactant or an anionic detergent;
b) lysing the host cell;
c) releasing the plasmid from the host cell and onto said medium; and
d) isolating said plasmid from said medium.
15. The method of claim 14 , wherein said solid medium further comprises uric acid or a urate salt.
16. The method of claim 14 , wherein said sample comprises host cells and wherein said host cells comprise eukaryotic cells.
17. The method of claim 14 , wherein said sample comprises host cells and wherein said host cells comprise prokaryotic cells.
18. The method of claim 12 , wherein said sample comprises host cells and wherein said host cells comprise eukaryotic cells.
19. The method of claim 12 , wherein said sample comprises host cells and wherein said host cells comprise prokaryotic cells.