Methods and compositions for arraying nucleic acids onto a solid support

• US 6,558,907 B2
• Filed: 05/16/2001
• Issued: 05/06/2003
• Est. Priority Date: 05/16/2001
• Status: Expired due to Fees

• Current Assignee: Corning Incorporated
• Original Assignee: Corning Incorporated
• Inventors: Koroulis, Melanie C., Pal, Santona
• Primary Examiner(s): Siew, Jeffrey
• Application Number:
  US09/859,160
• Publication Number:
  US 20020187476A1
• Time in Patent Office: 720 Days
• Field of Search: 435/6, 435/7.1, 435/91.1, 435/91.2, 435/287.2, 536/22.1, 536/23.1, 536/24-- 2
• US Class Current: 506/16
• CPC Class Codes: C12Q 1/6813 Hybridisation assays
• CPC Class Codes: C12Q 1/6837 using probe arrays or probe chips C12Q1/6874 takes precedence
• CPC Class Codes: C12Q 2527/137 Concentration of a component of medium
• CPC Class Codes: C12Q 2565/501 being an array of oligonucleotides

Abstract

The present invention provides a method for depositing a nucleic acid on a solid support. The method comprises contacting a solid support with a solution of nucleic acid, the solution comprising about 30% to about 80% dimethylsulfoxide (DMSO) by volume, sodium chloride and sodium citrate salt containing buffer (SSC) at a final concentration of from about 0.1× (15 mM sodium chloride+1.5 mM sodium citrate) to about 0.8× (120 mM sodium chloride+12 mM sodium citrate). The composition includes a nucleic acid at a concentration ranging from 0.01 mg/ml to 0.50 mg/ml. Preferably, the solution comprises about 40% to about 80% DMSO by volume and SSC at a final concentration from about 0.1× to about 0.5×. More preferably, the solution comprises about 40% to about 60% DMSO by volume and SSC at final concentration from about 0.25× to about 0.5×. Most preferably, the solution comprises about 50% DMSO by volume and SSC at a final concentration of about 0.25×. The nucleic acid is preferably a double stranded DNA or an oligonucleotide.

Claims

1. A medium for suspending a solution of nucleic acid, the medium having a composition comprising:

dimethylsulfoxide (DMSO) in a range from about 30% to about 80% by volume, sodium chloride/sodium citrate salt containing buffer (SSC) at a final concentration of from about 0.1×

(15 mM sodium chloride+1.5 mM sodium citrate) to about 0.8×

in salts (120 mM sodium chloride+12 mM sodium citrate), water, and nucleic acid, wherein said nucleic acid denatures to provide for more favorable hybridization.

2. The medium according to claim 1, wherein said solution induces said nucleic acid to show increased fluorescent signal when hybridized.

3. The medium according to claim 1, wherein said medium comprises about 40% to about 80% DMSO by volume and SSC at a final concentration from about 0.1×

to about 0.5×

.

4. The medium according to claim 1, wherein said medium comprises about 40% to about 60% DMSO by volume and SSC at a final concentration from about 0.25×

to about 0.5×

.

5. The medium according to claim 1, wherein said medium comprises about 50% DMSO by volume and SSC at a final concentration from about 0.1×

to about 0.5×

.

6. The medium according to claim 1, wherein said composition may also include EDTA in a final concentration between 0 and 4 mM.

7. The medium according to claim 1, wherein said nucleic acid is at a concentration ranging from about 0.01 mg/ml to about 0.5 mg/ml.

8. The medium according to claim 1, wherein the nucleic acid is a double stranded DNA, RNA, or an oligonucleotide.

9. The medium according to claim 1, wherein said composition enables long-term storage and preserves integrity of nucleic acid without instability by precipitation or aggregation of said nucleic acid.

10. The medium according to claim 9, wherein said composition enables long term storage of over 1 month.

11. The medium according to claim 1, wherein said medium absorbs moisture from air to overcome a net loss of solvent due to evaporation of said water component.

12. The medium according to claim 1, wherein said nucleic acid may be suspended in said medium for at least 5 days prior to depositing on a solid support.

13. A method for depositing nucleic acid on a solid support, said method comprising:

providing a medium according to that of claim 1, wherein said nucleic acid shows increased fluorescent signal when hybridized; and

contacting said medium to said solid support.

14. The method according to claim 13, wherein the contacting step further comprises immersing a tip of a pin into said medium;

removing said tip from said medium with said medium adhered to said tip; and

transferring said solution to said solid support.

15. The method according to claim 13, wherein said contacting step is repeated a plurality of times to provide an array of nucleic acid.

16. The method according to claim 13, wherein said nucleic acid is suspended in said medium for at least 5 days prior to contacting with said solid support.

17. The method according to claim 13, wherein said medium comprises about 40% to about 80% DMSO by volume and SSC at a final concentration from about 0.1×

to about 0.5×

.

18. The method according to claim 13, wherein said medium comprises about 40% to about 60% DMSO by volume and SSC at a final concentration from about 0.25'"'"' to about 0.5×

.

19. The method according to claim 13, wherein said medium comprises about 50% DMSO by volume and SSC at a final concentration from about 0.1×

to about 0.5×

.

20. The method according to claim 13, wherein said nucleic acid is at a concentration ranging from about 0.01 mg/ml to about 0.5 mg/ml.

21. The method according to claim 13, wherein the nucleic acid is a double stranded DNA, RNA, or an oligonucleotide.

22. The method according to claim 13, wherein said solid support is a membrane or glass substrate.

23. The method according to claim 22, wherein said glass substrate is a planar solid glass surface or three-dimensional porous glass surface.

24. The method according to claim 23, wherein said glass substrate comprises an aminated surface.

25. The method according to claim 24, wherein said aminated surface is coated with an aminating agent comprising either gama-aminopropyl-silane or polylysine.