STABILIZATION OF NUCLEIC ACIDS ON SOLID SUPPORTS

• US 20090053704A1
• Filed: 08/24/2007
• Published: 02/26/2009
• Est. Priority Date: 08/24/2007
• Status: Active Application

• Current Assignee: Agilent Technologies, Inc.
• Original Assignee: Agilent Technologies, Inc.
• Inventors: Braman, Jeffrey C., Basehore, Lee Scott, NOVORADOVSKAYA, Natalia
• Application Number:
  US11/844,578
• Publication Number:
  US 20090053704A1
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: C12Q 1/6806 Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 2527/125 Specific component of sample, medium or buffer

Abstract

The present invention provides methods, compositions, and kits for the storage and stabilization of biological molecules. The methods comprise applying Tris(2-carboxyethyl)phosphine (TCEP) to at least one biological molecule bound to a solid substrate and storing in an organic solvent. Preferably, the biological molecules are nucleic acids. Compositions and kits for performing the process according to the invention are also provided.

Claims

1. A method for stabilizing a biological molecule bound to a solid substrate, said method comprising:

contacting the biological molecule with at least one reducing agent; and

contacting the biological molecule with at least one organic solvents,

wherein said method steps results in stabilization of the biological molecule bound to the solid substrate.

2. The method of claim 1, further comprising removing some or all of the reducing agent(s).

3. The method of claim 1, further comprising storing the biological molecule in the presence of the organic solvent.

4. The method of claim 3, wherein the biological molecule is stored for period of time between about 1 hour to about 12 months.

5. The method of claim 1, further comprising causing the biological molecule to become unbound from the solid substrate.

6. The method of claim 1, wherein the biological molecule is a nucleic acid molecule.

7. The method of claim 6, wherein the nucleic acid is RNA.

8. The method of claim 1, wherein the organic solvent is ethanol, acetonitrile, acetone, tetrahydrofuran, 1,3-dioxolane, morpholine, tetraglyme, dimethyl sulfoxide, sulfolane, or a mixture of two or more of these.

9. A method for stabilizing a biological molecule bound to a solid substrate, said method comprising:

contacting the biological molecule with Tris(2-carboxyethyl)phosphine (TCEP); and

contacting the biological molecule with at least one organic solvents,

wherein said method steps results in stabilization of the biological molecule bound to the solid substrate.

10. The method of claim 9, further comprising removing some or all of the TCEP.

11. The method of claim 9, wherein the biological molecule is not contacted with (hydroxymethyl)aminomethane (Tris) or Tris-HCl.

12. The method of claim 9, further comprising storing the biological molecule in the presence of the organic solvent.

13. The method of claim 12, wherein the biological molecule is stored for a period of time between about 1 hour to about 12 months.

14. The method of claim 9, further comprising causing the biological molecule to become unbound from the solid substrate.

15. The method of claim 9, wherein the biological molecule is a nucleic acid molecule.

16. The method of claim 15, wherein the nucleic acid is RNA.

17. The method of claim 9, wherein the organic solvent is ethanol, acetonitrile, acetone, tetrahydrofuran, 1,3-dioxolane, morpholine, tetraglyme, dimethyl sulfoxide, sulfolane, or a mixture of two or more of these.

18. The method of claim 9, wherein the TCEP is provided in a composition having a pH of about 4 to about 8.

19. The method of claim 18, wherein the composition comprises 0.01 mM to 100 mM TCEP.

20. The method of claim 9, wherein contacting with at least one organic solvent creates a composition in which the organic solvent is at a final concentration of about 50% to 100% of the liquid in the composition.

21. A composition comprising:

a biological molecule bound to a solid support;

TCEP; and

at least one organic solvent,

wherein the concentrations of TCEP and organic solvent are sufficient to allow continued binding of the biological molecule to the solid substrate and to allow stabilization of the biological molecule at temperatures above 40°

C.

22. The composition of claim 21, wherein the composition comprises 0.01 mM to 100 mM TCEP and the pH of the composition is from about 4 to about 8.

23. The composition of claim 21, wherein the biological molecule is a nucleic acid.

24. The composition of claim 21, wherein the nucleic acid is RNA.

25. A stabilized RNA molecule prepared by:

binding the RNA to a solid support;

contacting the RNA with TCEP; and

contacting the RNA with at least one organic solvent.

26. The stabilized RNA of claim 25, wherein the RNA can be stored for at least 1 hour without appreciable degradation.

27. The stabilized RNA of claim 25, wherein the RNA can be stored for at least 30 minutes at a temperature greater than 0°

C.

28. The stabilized RNA of claim 25, wherein the RNA is bound to the solid support.

29. The stabilized RNA of claim 25, wherein the RNA is maintained in the presence of the one or more organic solvents.

30. (canceled)

31. (canceled)