Hybridization compositions and methods
The invention provides methods and compositions for hybridizing at least one molecule to a target. The invention may, for example, utilize a of cyclic and/or non-cyclic solvent that is non-toxic and may eliminate or reduce the amount of formamide in the hybridization composition.
1. A hybridization composition comprising at least one nucleic acid sequence, at least one solvent in an amount effective to denature double-stranded nucleotide sequences, and a hybridization solution, wherein the solvent is chosen from tetrahydrothiophene 1-oxide, valerolactam, 1,3-dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone, N,N-dimethyl-acetamide and isobutyramide.
2. The hybridization composition according to claim 1 , wherein the solvent is a non-cyclic solvent.
3. The hybridization composition according to claim 1 , wherein the solvent is chosen from N,N-dimethyl-acetamide, isobutyramide, and 1,3-dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone.
4. The hybridization composition according to claim 1 , wherein the concentration of solvent is about 1% to 95% (v/v).
5. The hybridization composition according to claim 1 , wherein the concentration of solvent is 5% to 10% (v/v).
6. The hybridization composition according to claim 1 , wherein the concentration of solvent is 10% to 20% (v/v).
7. The hybridization composition according to claim 1 , wherein the concentration of solvent is 20% to 30% (v/v).
8. The hybridization composition according to claim 1 , wherein the solvent is non-toxic.
9. The hybridization composition according to claim 8 , wherein the composition contains less than 10% (v/v) formamide.
10. The hybridization composition according to claim 9 , wherein the composition contains less than 2% (v/v) formamide.
11. The hybridization composition according to claim 10 , wherein the composition contains less than 1% (v/v) formamide.
12. The hybridization composition according to claim 1 , further comprising at least one additional component selected from the group consisting of: buffering agents, salts, accelerating agents, chelating agents, detergents, and blocking agents.
13. The hybridization composition according to claim 12 ,
wherein the at least one additional component is an accelerating agent, a salt, and/or a buffering agent; and
wherein the accelerating agent is dextran sulfate, the salt is NaCl, or the buffering agent is phosphate buffer.
14. The hybridization composition according to claim 13 , wherein:
the NaCl is present at a concentration of up to 1200 mM, and/or
the phosphate buffer is present at a concentration of up to 50 mM.
15. The hybridization composition according to claim 14 , wherein:
the NaCl is present at a concentration of 300 mM to 600 mM, and/or
the phosphate buffer is present at a concentration of 5 mM to 20 mM.
16. The hybridization composition according to claim 12 , wherein the accelerating agent is selected from the group consisting of: formamide, glycerol, propylene glycol, 1,2-propanediol, diethylene glycol, ethylene glycol, glycol, and 1,3 propanediol, and the buffering agent is citric acid buffer.
17. The hybridization composition according to claim 16 , wherein:
the formamide is present at a concentration of 0.1-5% (v/v),
the glycerol, propylene glycol, 1,2-propanediol, diethylene glycol, ethylene glycol, glycol, or 1,3 propanediol are present at a concentration of 0.1% to 10% (v/v), and
the citric acid buffer is present at a concentration of 1 mM to 50 mM.
18. The hybridization composition according to claim 1 , comprising 40% (vN) of the solvent, dextran sulfate, 300 mM NaCl, and 5 mM phosphate buffer.
19. The hybridization composition according to claim 1 , comprising 15% (vN/V) of the solvent, dextran sulfate, 600 mM NaCl, and 10 mM phosphate buffer.
20. The hybridization composition according to claim 1 , comprising 15% (vN) of the solvent, dextran sulfate, 600 mM NaCl, and 10 mM citric acid buffer pH 6.2.