Nucleic acid preservation solution and methods of manufacture and use
Disclosed is nucleic acid preserving compositions and methods of manufacturing and using the same. Compositions include a carrier, a chaotropic agent, a buffering agent, a chelating agent, a surfactant, an alcohol, an acid, and a mucolytic agent. Compositions as aqueous solutions can include water as a carrier. Preferred embodiments include water, guanidine thiocyanate, Tris, EDTA, SLS, SDA 3C, HCl, and N-acetyl-L-cysteine. Some embodiments include a colored dye as a visual indicator. Methods of manufacturing include combining the components into a mixture, such as an aqueous solution. Methods of use include providing a biological sample that includes nucleic acid and contacting the biological sample with the composition. Kits include the composition disposed in a portion of a biological sample collection apparatus.
1. A nucleic acid preservation composition, comprising:
an aqueous carrier;
about 20% to about 50%, w/w, of a chaotropic agent;
about 0.1% to about 5%, w/w, of a buffering agent;
about 0.05% to about 2.5%, w/w, of a chelating agent comprising;
about 0.01% to about 5%, w/w, of a detergent or a surfactant;
about 5% to about 25%, w/w, of an alcohol, wherein the alcohol comprises a mixture of about 95% ethanol, v/v, and about 5% isopropanol, v/v; and
0.005% to about 0.5%, w/w, of a mucolytic agent or a reducing agent, wherein the mucolytic agent or reducing agent comprises N-acetyl-L-cysteine (NAC),
the composition having a pH of 6.5-9.5.
2. The composition of claim 1 , wherein the composition has a pH of about 7 to about 9.
3. The composition of claim 1 , wherein:
the chaotropic agent comprises guanidine thiocyanate;
the buffering agent comprises tris(hydroxymethyl)aminomethane (Tris);
the chelating agent comprises ethylenediaminetetraacetic acid (EDTA) or EDTA disodium (salt) dihydrate; and
the detergent or surfactant comprises sodium lauroyl sarcosinate (SLS).
4. The composition of claim 1 , wherein the composition is substantially free or devoid of antimicrobial agent(s), bactericidal agent(s), and/or bacteriostatic agent(s) other than the chaotropic agent, the detergent or surfactant, the alcohol, and the mucolytic agent or reducing agent.
5. The composition of claim 1 , wherein the composition is substantially free or devoid of protease(s).
6. The composition of claim 1 , wherein the composition is substantially free or devoid of inhibitor(s) of ribonuclease other than the chaotropic agent, the surfactant, the alcohol, and the mucolytic agent.
7. A nucleic acid preservation kit, comprising:
a sample collection apparatus comprising a solution compartment; and
the nucleic acid preservation composition of claim 1 disposed in the solution compartment.
8. A method of preserving nucleic acid in a biological sample, the method comprising:
obtaining or providing a biological sample containing nucleic acid, the biological sample optionally comprising saliva; and
contacting the biological sample with the nucleic acid preservation composition of claim 1 .
9. A nucleic acid preservation composition, comprising:
guanidine thiocyanate;
tris(hydroxymethyl)aminomethane (Tris);
ethylenediaminetetraacetic acid (EDTA) or EDTA disodium (salt) dihydrate;
sodium lauroyl sarcosinate (SLS);
an alcohol, wherein the alcohol comprises a mixture of about 95% ethanol, v/v, and about 5% isopropanol, v/v; and
N-acetyl-L-cysteine (NAC).
10. The composition of claim 9 , wherein the composition has a pH of 6.5-9.5.
11. The composition of claim 9 , wherein the composition has a pH of about 7 to about 9.
12. The composition of claim 9 , wherein the composition is substantially free or devoid of antimicrobial agent(s), bactericidal agent(s), and/or bacteriostatic agent(s) other than the chaotropic agent, the detergent or surfactant, the alcohol, and the mucolytic agent or reducing agent.
13. The composition of claim 9 , wherein the composition is substantially free or devoid of protease(s).
14. The composition of claim 9 , wherein the composition is substantially free or devoid of inhibitor(s) of ribonuclease other than the chaotropic agent, the surfactant, the alcohol, and the mucolytic agent.
15. A nucleic acid preservation kit, comprising:
a sample collection apparatus comprising a solution compartment; and
the nucleic acid preservation composition of claim 9 disposed in the solution compartment.
16. A method of preserving nucleic acid in a biological sample, the method comprising:
obtaining or providing a biological sample containing nucleic acid, the biological sample optionally comprising saliva; and
contacting the biological sample with the nucleic acid preservation composition of claim 9 .
17. A nucleic acid preservation composition, comprising:
an aqueous carrier;
a chaotropic agent;
a buffering agent;
a chelating agent;
a detergent or a surfactant;
an alcohol, wherein the alcohol comprises a mixture of about 95% ethanol, v/v, and about 5% isopropanol, v/v; and
N-acetyl-L-cysteine (NAC).
18. The nucleic acid preservation composition of claim 17 , wherein the composition has a pH of about 7 to about 9.
19. The nucleic acid preservation composition of claim 18 , wherein:
the composition is substantially free or devoid of antimicrobial agent(s), bactericidal agent(s), and/or bacteriostatic agent(s) other than the chaotropic agent, the detergent or surfactant, the alcohol, and the mucolytic agent or reducing agent;
the composition is substantially free or devoid of protease(s); and
the composition is substantially free or devoid of inhibitor(s) of ribonuclease other than the chaotropic agent, the surfactant, the alcohol, and the mucolytic agent.
20. The nucleic acid preservation composition of claim 19 , wherein:
the chaotropic agent comprises guanidine thiocyanate;
the buffering agent comprises tris(hydroxymethyl)aminomethane (Tris);
the chelating agent comprises ethylenediaminetetraacetic acid (EDTA) or EDTA disodium (salt) dihydrate; and
the detergent or surfactant comprises sodium lauroyl sarcosinate (SLS).