STABILIZATION OF NUCLEIC ACIDS ON SOLID SUPPORTS

• US 20110230653A1
• Filed: 05/27/2011
• Published: 09/22/2011
• Est. Priority Date: 08/24/2007
• Status: Abandoned Application

• Current Assignee: Agilent Technologies, Inc.
• Original Assignee: Agilent Technologies, Inc.
• Inventors: BASEHORE, Lee Scott, NOVORADOVSKAYA, Natalia, BRAMAN, Jeffrey C.
• Application Number:
  US13/117,935
• Publication Number:
  US 20110230653A1
• Time in Patent Office: Days
• US Class Current: 536/23.1
• 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 and storing a nucleic acid bound to a solid substrate, said method comprising:

contacting the nucleic acid with at least one reducing agent;

contacting the nucleic acid with at least one organic solvent; and

storing the nucleic acid bound to the solid substrate at a temperature of about 4°

C. or higher for at least 1 hour,

wherein said method steps results in stabilization of the nucleic acid bound to the solid substrate for at least 1 hour at a temperature about 4°

C. or higher, and

wherein the solid substrate is not alumina.

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

3. The method of claim 1, wherein the method comprises storing the nucleic acid in the presence of the organic solvent.

4. The method of claim 3, wherein the nucleic acid is stored for period of time between about 1 day to about 12 months.

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

6. The method of claim 1, wherein the nucleic acid is RNA.

7. The method of claim 1, wherein the nucleic acid is DNA.

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 and storing a nucleic acid bound to a solid substrate, said method comprising:

contacting the nucleic acid with Tris(2-carboxyethyl)phosphine (TCEP);

contacting the nucleic acid with at least one organic solvent; and

storing the nucleic acid bound to the solid substrate at a temperature of about 4°

C. or higher for at least 1 hour,

wherein said method steps result in stabilization of the nucleic acid bound to the solid substrate for at least 1 hour at a temperature of about 4°

C. or higher, and

wherein the solid substrate is not alumina.

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

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

12. The method of claim 9, wherein the method comprises storing the nucleic acid in the presence of the organic solvent.

13. The method of claim 12, wherein the nucleic acid 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 nucleic acid to become unbound from the solid substrate.

15. The method of claim 9, wherein the nucleic acid is RNA.

16. The method of claim 9, wherein the nucleic acid is DNA.

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.