Polynucleotides, reagents, and methods for nucleic acid hybridization
Provided herein are compositions, methods and systems relating to libraries of polynucleotides such that the libraries allow for accurate and efficient hybridization after binding to target sequences. Further provided herein are probes, blockers, additives, buffers, and methods that result in improved hybridization. Such compositions and methods are useful for improvement of Next Generation Sequencing applications, such as reducing off-target binding or reducing workflow times.
1. An aqueous composition for polynucleotide hybridization comprising:
at least one solvent, wherein the at least one solvent comprises dimethyl sulfoxide (DMSO) at 10-30% (v/v);
at least one surfactant; and
at least one thickening agent.
2. The composition of claim 1 , wherein the at least one solvent further comprises DMF, DMAc, or HEMPA.
3. The composition of claim 1 , wherein the at least one solvent further comprises acetone, methyl ethyl ketone, THF, or diethyl ether.
4. The composition of claim 1 , wherein the at least one surfactant comprises SDS, CTAB, tergitol, or sodium lauryl sulfate.
5. The composition of claim 1 , wherein the at least one surfactant comprises a trialkylammonium salt.
6. The composition of claim 1 , wherein the at least one thickening agent comprises cellulose, or starch.
7. The composition of claim 1 , wherein the at least one thickening agent comprises dextran sulfate, hydroxymethylcellulose, hydroxyethylcellulose, polyethylene glycol, or ficoll.
8. The composition of claim 1 , wherein the at least one thickening agent is 10-40% (w/v).
9. The composition of claim 1 , wherein the at least one surfactant is 0.001-0.1% (w/v).
10. The composition of claim 4 , wherein the at least one surfactant is 0.001-0.1% (w/v).
11. The composition of claim 5 , wherein the at least one surfactant is 0.001-0.1% (w/v).
12. The composition of claim 6 , wherein the at least one thickening agent is 10-40% (w/v).
13. The composition of claim 7 , wherein the at least one thickening agent is 10-40% (w/v).
14. The composition of claim 1 , wherein a hybridization temperature of the composition is at least 50° C.
15. The composition of claim 1 , wherein a hybridization temperature of the composition is at least 65° C.
16. The composition of claim 1 , wherein the aqueous composition comprises a plurality of polynucleotides.
17. The composition of claim 16 , wherein the plurality of polynucleotides are in solution.