Methods and compositions 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 template nucleic acids, wherein each template nucleic acid comprises a first end, and a second end capable of hybridizing to SEQ ID NO:05, wherein a plurality of the template nucleic acids in the population are different from each other.
2. The population of claim 1 , wherein the second end comprises SEQ ID NO:04.
3. The population of claim 1 , wherein the first end is capable of hybridizing to SEQ ID NO:03.
4. The population of claim 1 , wherein the first end comprises SEQ ID NO:11.
5. The population of claim 1 , wherein a remainder polynucleotide is disposed between the first end and the second end.
6. The population of claim 1 , wherein the template nucleic acids have a length greater than 300 nucleotides.
7. The population of claim 1 , wherein the template nucleic acids comprise genomic DNA.
8. The population claim 1 , wherein the first end or second end is capable of hybridizing at 5×SSC and 40° C.
9. A composition comprising the population of template nucleic acids of claim 1 hybridized to a plurality of first oligonucleotides via the first ends of the template nucleic acids, wherein the first oligonucleotides are immobilized on a substrate.
10. The composition of claim 9 , wherein the first ends are capable of hybridizing to SEQ ID NO:03.
11. The composition of claim 9 , wherein a plurality of the first ends comprise SEQ ID NO:11.
12. The composition of claim 9 , wherein the first oligonucleotides comprise SEQ ID NO:03.
13. The composition of claim 9 , wherein a plurality of the second ends of the template nucleic acids comprise SEQ ID NO:04.
14. The composition of claim 9 , further comprising a plurality of second oligonucleotides immobilized on a substrate.
15. The composition of claim 14 , wherein the plurality of second oligonucleotides comprise SEQ ID NO:04.
16. The composition of claim 9 , wherein the immobilized first oligonucleotides each comprise a polyT spacer comprising from 2 to 10 thymine nucleotides, and wherein the polyT spacer is at the 5′ end of the immobilized first oligonucleotides.
17. The composition of claim 9 , wherein the plurality of first oligonucleotides is covalently attached to a hydrogel on the substrate.
18. A method of amplifying a population of template nucleic acids comprising:
hybridizing the population of template nucleic acids of claim 1 to a plurality of first oligonucleotides via the first ends of the template nucleic acids, wherein the plurality of first oligonucleotides are immobilized on a substrate;
extending the first oligonucleotides;
hybridizing the extended first oligonucleotides to a plurality of second oligonucleotides immobilized on the substrate; and
extending the hybridized second oligonucleotides to amplify the template nucleic acids.
19. The method of claim 18 , further comprising removing the population of template nucleic acids from the extended first oligonucleotides.
20. The method of claim 18 , further comprising sequencing the extended first oligonucleotides or the extended second oligonucleotides.