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 capable of hybridizing to SEQ ID NO:03, and a second end, wherein the template nucleic acids are different from each other and comprise genomic DNA.
2. The population of claim 1 , wherein the first end comprises SEQ ID NO:11.
3. The population of claim 1 , wherein the second end is capable of hybridizing to SEQ ID NO:05.
4. The population of claim 1 , wherein the second end comprises SEQ ID NO: 4.
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 claim 1 , wherein the first end is capable of hybridizing at 5×SSC and 40° C.
8. A composition comprising: a population of template nucleic acids that are different from one another, and wherein each template nucleic acid comprises a first end capable of hybridizing to SEQ ID NO:03 and a second end, wherein the population of template nucleic acids is 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.
9. The composition of claim 8 , wherein the first oligonucleotides comprise SEQ ID NO:03.
10. The composition of claim 8 , wherein the second ends of the template nucleic acids are capable of hybridizing to SEQ ID NO:05.
11. The composition of claim 10 , wherein the second ends of the template nucleic acids comprise SEQ ID NO: 4.
12. The composition of claim 8 , further comprising a plurality of second oligonucleotides immobilized on a substrate.
13. The composition of claim 12 , wherein the plurality of second oligonucleotides comprise SEQ ID NO:04.
14. The composition of claim 8 , wherein the immobilized first oligonucleotides each comprise a polyT spacer comprising from 2 to 10 thymine nucleotides, wherein the polyT spacer is at the 5′ end of the immobilized first oligonucleotides.
15. The composition of claim 8 , wherein the plurality of first oligonucleotides is covalently attached to a hydrogel on the substrate.
16. 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.
17. The method of claim 16 , wherein the plurality of first oligonucleotides comprise SEQ ID NO:03.
18. The method of claim 16 , wherein the second ends of the template nucleic acids are capable of hybridizing to SEQ ID NO:05.
19. The method of claim 18 , wherein the second ends comprise SEQ ID NO: 4.
20. The method of claim 16 , wherein the plurality of second oligonucleotides comprise SEQ ID NO:04.
21. The method of claim 16 , further comprising removing the population of template nucleic acids from the extended first oligonucleotides.
22. The method of claim 16 , further comprising sequencing the extended first oligonucleotides or the extended second oligonucleotides.