AMPLIFICATION OLIGONUCLEOTIDES
Disclosed herein, inter alia, are polynucleotides, supports, kits, and methods of use thereof for amplifying, immobilizing, and sequencing polynucleotides.
1 .- 64 . (canceled)
65 . A circular polynucleotide comprising
a first cleavable site;
a first sequencing primer sequence, wherein the first sequencing primer sequence comprises SEQ ID NO:152;
a template polynucleotide sequence; and
a second cleavable site.
66 . The circular polynucleotide of claim 65 , further comprising a second sequencing primer binding sequence; wherein the second sequencing primer binding sequence comprises SEQ ID NO:178.
67 . The circular polynucleotide of claim 65 , further comprising a first platform primer sequence and a second platform primer sequence.
68 . The circular polynucleotide of claim 67 , wherein the first platform primer sequence comprises SEQ ID NO:92.
69 . The circular polynucleotide of claim 67 , wherein the second platform primer sequence comprises 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.
70 . The circular polynucleotide of claim 67 , wherein the second platform primer sequence comprises SEQ ID NO:104.
71 . The circular polynucleotide of claim 67 , wherein the second platform primer sequence comprises SEQ ID NO:106.
72 . The circular polynucleotide of claim 67 , wherein the second platform primer sequence comprises SEQ ID NO:4.
73 . The circular polynucleotide of claim 65 , wherein the template polynucleotide sequence comprises genomic DNA.
74 . The circular polynucleotide of claim 65 , wherein the template polynucleotide sequence comprises a cancer associated gene, or fragment thereof.
75 . The circular polynucleotide of claim 65 , wherein the first and the second cleavable site independently comprise a diol linker, disulfide linker, or a photocleavable linker.
76 . The circular polynucleotide of claim 65 , wherein the first and the second cleavable site independently comprise an abasic site, uracil nucleotide, deoxy-8-oxo-guanine nucleotide, methylated nucleotide, or a ribonucleotide.
77 . The circular polynucleotide of claim 65 , wherein the first and the second cleavable site each comprise an abasic site.
78 . The circular polynucleotide of claim 65 , wherein the first and the second cleavable site each comprise a uracil nucleotide.
79 . The circular polynucleotide of claim 65 , wherein the first and the second cleavable site each comprise a ribonucleotide.
80 . The circular polynucleotide of claim 65 , wherein the first and the second cleavable site independently comprise a sequence specifically recognized by an endonuclease enzyme or an exonuclease enzyme.
81 . The circular polynucleotide of claim 67 , further comprising a first index sequence and a second index sequence.
82 . The circular polynucleotide of claim 81 , wherein the first index sequence and the second index sequence independently comprise 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 nucleotides.
83 . The circular polynucleotide of claim 81 , wherein the first index sequence and the second index sequence independently comprise 6 to 12 nucleotides.
84 . The circular polynucleotide of claim 81 , wherein the first index sequence is between the first platform primer sequence and the first sequencing primer sequence.