Amplification oligonucleotides
Disclosed herein, inter alia, are polynucleotides, supports, kits, and methods of use thereof for amplifying, immobilizing, and sequencing polynucleotides.
1. A plurality of nucleic acid molecules, wherein each nucleic acid molecule comprises a first end, a target sequence, and a second end, wherein the first end comprises SEQ ID NO:92, and wherein each target sequence of said plurality of nucleic acid molecules is different from each other.
2. The plurality of claim 1 , wherein the second end comprises a sequence comprising SEQ ID NO:104, SEQ ID NO:4, SEQ ID NO:102, SEQ ID NO:106, SEQ ID NO:108, SEQ ID NO:134, SEQ ID NO:136, SEQ ID NO:138, or SEQ ID NO:140.
3. The plurality of claim 2 , wherein the second end comprises SEQ ID NO:4.
4. The plurality of claim 2 , wherein the second end comprises SEQ ID NO:136.
5. The plurality of claim 1 , wherein the target sequence comprises genomic DNA.
6. The plurality of claim 1 , wherein the target sequence comprises a cancer associated gene, or fragment thereof.
7. The plurality of claim 1 , wherein each nucleic acid molecule comprises 50 to 1000 nucleotides.
8. The plurality of claim 1 , wherein each nucleic acid molecule comprises 100 to 500 nucleotides.
9. The plurality of claim 1 , wherein each nucleic acid molecule consists of 50 to 1000 nucleotides.
10. The plurality of claim 1 , wherein one of said first end or said second end comprise a hairpin adapter.
11. The plurality of claim 1 , wherein said first end and said second end each comprise a hairpin adapter.
12. The plurality of claim 1 , wherein one of said first end or said second end comprise a Y polynucleotide adapter.
13. The plurality of claim 1 , wherein both said first end and said second end comprise a Y polynucleotide adapter.
14. The plurality of claim 1 , wherein said plurality of nucleic acid molecules comprises single-stranded DNA or double-stranded DNA.
15. The plurality of claim 1 , wherein each nucleic acid molecule comprises, from 5′ to 3′, said first end, said target sequence, and said second end.
16. The plurality of claim 1 , wherein the second end comprises SEQ ID NO:104.
17. The plurality of claim 1 , wherein one or more of the nucleic acid molecules are immobilized to a solid support.
18. The plurality of claim 17 , wherein said solid support further comprises one or more complements of said nucleic acid molecules.
19. The plurality of claim 17 , wherein said solid support further comprises a polymer, resist, and/or hydrogel.
20. The plurality of claim 17 , wherein said nucleic acid molecules are covalently attached to a polymer on the solid support via a linker.
21. The plurality of claim 20 , wherein the linker comprises 8 to 16 thymine nucleotides, and wherein the linker is at a 5′ end of the immobilized nucleic acid molecules.
22. The plurality of claim 20 , wherein the linker comprises a cleavable site.
23. The plurality of claim 22 , wherein the cleavable site comprises a deoxyuracil triphosphate (dUTP), diol linker, disulfide linker, photocleavable linker, abasic site, deoxy-8-Oxo-guanine triphosphate (d-8-oxoG), methylated nucleotide, ribonucleotide, or a sequence containing a modified or unmodified nucleotide that is specifically recognized by a cleaving agent.
24. The plurality of claim 17 , wherein said one or more nucleic acid molecules further comprise a first oligonucleotide hybridized to the first end, and wherein said first oligonucleotide is attached to said solid support.
25. The plurality of claim 17 , wherein said one or more nucleic acid molecules further comprise a second oligonucleotide hybridized to the second end, and wherein said second oligonucleotide is attached to said solid support.
26. The plurality of claim 17 , further comprising one or more complements of said nucleic acid molecules immobilized to said solid support.
27. The plurality of claim 26 , wherein said one or more immobilized complements of said nucleic acid molecules are hybridized to said immobilized nucleic acid molecules.
28. The plurality of claim 17 , wherein said one or more immobilized nucleic acid molecules is hybridized to a sequencing primer, and wherein said sequencing primer comprises a labeled nucleotide at a 3′ end.