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 human nucleic acid preservation composition, comprising:
39.53%-48.31% guanidine thiocyanate, w/w;
2.39%-2.92% tris(hydroxymethyl)aminomethane (Tris), w/w;
0.73%-0.89% ethylenediaminetetraacetic acid (EDTA) or EDTA disodium (salt) dihydrate, w/w;
0.251%-0.307% sodium lauroyl sarcosinate (SLS), w/w;
15.96%-19.50% alcohol, w/w, the alcohol comprising ethanol;
0.084%-0.102% N-acetyl-L-cysteine (NAC), w/w;
hydrochloric acid qs to pH 7.5-8.5; and
an aqueous carrier comprising filtered, purified, distilled, and/or deionized water.
2. The composition of claim 1 further comprising a dye or coloring agent.
3. The composition of claim 1 , wherein the composition comprises:
43.48%-44.36%, w/w, of the guanidine thiocyanate;
2.62%-2.68%, w/w, of the Tris;
0.80%-0.82%, w/w, of the EDTA or EDTA disodium (salt) dihydrate;
0.276%-0.282%, w/w, of the SLS;
17.55%-17.91%, w/w, of the alcohol;
0.092%-0.094%, w/w, of the NAC; and
the hydrochloric acid qs to pH 7.8-8.2.
4. The composition of claim 1 , wherein the composition is substantially free or devoid of mucolytic agent(s) and/or reducing agent(s) other than N-acetyl-L-cysteine.
5. The composition of claim 1 , wherein the composition comprises:
41.72%-46.12%, w/w, of the guanidine thiocyanate;
2.52%-2.78%, w/w, of the Tris;
0.77%-0.85%, w/w, of the EDTA or EDTA disodium (salt) dihydrate;
0.265%-0.293%, w/w, of the SLS;
16.84%-18.62%, w/w, of the alcohol;
0.088%-0.098%, w/w, of the NAC; and
The acid qs to pH 7.6-8.4.
6. 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 guanidine thiocyanate, SLS, alcohol, and NAC.
7. The composition of claim 1 , wherein the composition is substantially free or devoid of inhibitor(s) of ribonuclease other than the guanidine thiocyanate, SLS, alcohol, and NAC.
8. The composition of claim 1 , wherein the composition is substantially free or devoid of protease(s).
9. The composition of claim 1 , wherein the alcohol comprises a mixture of about 95% ethanol, v/v, and about 5% isopropanol, v/v.
10. A nucleic acid preservation composition, comprising:
a carrier;
20-50%, w/w, of a chaotropic agent;
1-5%, w/w, of a buffering agent;
0.05-2.5%, w/w, of a chelating agent;
0.05-2.5%, w/w, of a surfactant;
5-25%, w/w, of alcohol;
an acid qs to pH 7.2-8.8; and
0.005-0.25%, w/w, of a mucolytic agent.
11. The composition of claim 10 , wherein the composition comprises:
39.53%-48.31%, w/w, of the chaotropic agent;
2.39%-2.92%, w/w, of the buffering agent;
0.73%-0.89%, w/w, of the chelating agent;
0.251%-0.307%, w/w, of the surfactant;
15.96%-19.50%, w/w, of the alcohol;
0.084%-0.102%, w/w, of the mucolytic agent; and
the acid qs to pH 7.5-8.5%.
12. The composition of claim 11 , 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 surfactant, the alcohol, and the mucolytic agent.
13. The composition of claim 11 , 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.
14. The composition of claim 11 , wherein the composition is substantially free or devoid of protease(s).
15. The composition of claim 10 , wherein the composition comprises:
43.48%-44.36%, w/w, of the chaotropic agent;
2.62%-2.68%, w/w, of the buffering agent;
0.80%-0.82%, w/w, of the chelating agent;
0.276%-0.282%, w/w, of the surfactant;
17.55%-17.91%, w/w, of the alcohol;
0.092%-0.094%, w/w, of the mucolytic agent; and
the acid qs to pH 7.8-8.2.
16. The composition of claim 15 , 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 surfactant, the alcohol, and the mucolytic agent.
17. The composition of claim 15 , 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.
18. The composition of claim 15 , wherein the composition is substantially free or devoid of protease(s).
19. The composition of claim 10 , wherein:
The carrier comprises filtered, purified, distilled, and/or deionized water;
the chaotropic agent comprises guanidine thiocyanate;
the buffering agent comprises tris(hydroxymethyl)aminomethane (Tris);
the chelating agent comprises ethylenediaminetetraacetic acid (EDTA)
the surfactant comprises sodium lauroyl sarcosinate (SLS);
the alcohol comprises the acid comprises hydrochloric acid (HCl); and
the mucolytic agent comprises N-Acetyl-L-cysteine (NAC).
20. The composition of claim 10 , furthering comprising a dye or coloring agent.
21. The composition of claim 10 , wherein the composition is substantially free or devoid of mucolytic agent(s) and/or reducing agent(s) other than N-acetyl-L-cysteine.
22. The composition of claim 10 , wherein the composition comprises:
41.72%-46.12%, w/w, of the chaotropic agent;
2.52%-2.78%, w/w, of the buffering agent;
0.77%-0.85%, w/w, of the chelating agent;
0.265%4293%, w/w, of the surfactant;
16.84%-18.62%, w/w, of the alcohol;
0.088%-0.098%, w/w, of the mucolytic agent; and
The acid qs to pH 7.6-8.4.
23. The composition of claim 10 , 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 surfactant, the alcohol, and the mucolytic agent.
24. The composition of claim 10 , 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.
25. The composition of claim 10 , wherein the composition is substantially free or devoid of protease(s).