Methods of localizing nucleic acids to arrays
Methods and compositions are disclosed relating to the localization of nucleic acids to arrays such as silane-free arrays, and of sequencing the nucleic acids localized thereby.
1. A population of polynucleotides comprising template nucleic acids having a first end capable of hybridizing to SEQ ID NO: 3, a second end capable of hybridizing to SEQ ID NO: 5 and a remainder polynucleotide disposed between the first end and the second end, wherein the template nucleic acids are different from each other.
2. The population of claim 1 , wherein the template nucleic acids comprise genomic DNA.
3. The population of claim 1 , further comprising amplified template nucleic acids.
4. The population of claim 1 , wherein the template nucleic acids comprise more than 300 base pairs of nucleotides.
5. The population of claim 1 , wherein the first end comprises SEQ ID NO: 11, and the second end comprises SEQ ID NO: 4.
6. A composition comprising:
a solid support comprising a surface;
a plurality of immobilized nucleic acids bound to the surface, wherein the immobilized nucleic acids each comprise a primer sequence selected from SEQ ID NO: 3 and SEQ ID NO:4; and
a population of template nucleic acids, wherein each template nucleic acid comprises a first end capable of hybridizing to SEQ ID NO: 3 and a second end capable of hybridizing to SEQ ID NO: 5, wherein at least one template nucleic acid is hybridized to an immobilized nucleic acid comprising SEQ ID NO: 3.
7. The composition of claim 6 , wherein each first end of each template nucleic acid is hybridized to an immobilized nucleic acid comprising SEQ ID NO: 3.
8. The composition of claim 6 , wherein the template nucleic acids are different from each other.
9. The composition of claim 6 , wherein the template nucleic acids comprise genomic DNA.
10. The composition of claim 6 , wherein the template nucleic acids comprise more than 300 base pairs of nucleotides.
11. The composition of claim 6 , wherein the first end comprises SEQ ID NO:11, and the second end comprises SEQ ID NO: 4.
12. The composition of claim 6 , wherein the immobilized nucleic acids each comprise a polyT spacer 5′ of the primer sequence comprising from 2 to 10 thymine nucleotides.
13. The composition of claim 6 , wherein the plurality of immobilized nucleic acids is covalently attached to a hydrogel on the surface.
14. The composition of claim 13 , wherein the hydrogel is the product of polymerizing a first comonomer with a second comonomer.
15. The composition of claim 14 , wherein the first comonomer is selected from the group consisting of acrylamide, methacrylamide, hydroxyethyl methacrylate and N-vinyl pyrrolidinone, and wherein the second comonomer is a functionalized comonomer selected from the group consisting of acrylamide, acrylate, methacrylate and methacrylamide.
16. A method of localizing a population of template nucleic acids to a solid support comprising:
providing a solid support comprising:
a surface, and
a plurality of nucleic acids immobilized on the surface, wherein the nucleic acids in the plurality each comprise a primer sequence selected from SEQ ID NO: 3 and SEQ ID NO:4; and
hybridizing a population of template nucleic acids having a first end capable of hybridizing to SEQ ID NO: 3 and a second end capable of hybridizing to SEQ ID NO: 5 to the plurality of nucleic acids comprising SEQ ID NO: 3 immobilized on the surface.
17. The method of claim 16 , further comprising extending the nucleic acids immobilized on the surface and hybridized to a template nucleic acid.
18. The method of claim 17 , further comprising determining the sequence of the extended nucleic acids.
19. The method of claim 16 , further comprising amplifying the population of nucleic acids comprising template nucleic acids.
20. The method of claim 16 , wherein the template nucleic acids are different from each other.
21. The method of claim 16 , wherein the template nucleic acids comprise genomic DNA.
22. The method of claim 16 , wherein the template nucleic acids comprise more than 300 base pairs of nucleotides.
23. The method of claim 16 , wherein the first end comprises SEQ ID NO:11, and the second end comprises SEQ ID NO: 4.
24. The method of claim 16 , wherein the nucleic acids in the plurality each comprise a polyT spacer 5′ of the primer sequence comprising from 2 to 10 thymine nucleotides.
25. The method of claim 16 , wherein the plurality of nucleic acids is covalently attached to a hydrogel immobilized on the surface.
26. The method of claim 25 , wherein the hydrogel is the product of polymerizing a first comonomer with a second comonomer.
27. The method of claim 26 , wherein the first comonomer is selected from the group consisting of acrylamide, methacrylamide, hydroxyethyl methacrylate and N-vinyl pyrrolidinone, and wherein the second comonomer is a functionalized comonomer selected from the group consisting of acrylamide, acrylate, methacrylate and methacrylamide.
28. The population claim 1 , wherein the first end and the second end are capable of hybridizing at 5×SSC and 40° C.
29. The composition of claim 6 , wherein the first end and the second end are capable of hybridizing at 5×SSC and 40° C.
30. The method of claim 16 , wherein the first end and the second end are capable of hybridization at 5×SSC and 40° C.