Nucleic acid ligation systems and methods
The present disclosure relates to systems and methods for nucleic acid ligation. In particular, the present disclosure provides oligonucleotide adaptors for use in nucleic acid ligation reactions.
1. A kit, comprising:
a plurality of double stranded nucleic acid adaptors, wherein each of said plurality of adaptors has a distinct single stranded overhang relative to each other adaptor, wherein said adaptor overhangs have random nucleotide sequences complementary to a plurality of single stranded nucleotide overhangs of one or more double stranded nucleic acid inserts; and a ligase.
2. The kit of claim 1 , wherein said plurality of double stranded nucleic acid adaptors comprises adaptor overhangs of 1 or 2 nucleotides.
3. The kit of claim 1 , wherein said adaptor overhangs have single nucleotide overhangs at the 3′ or 5′ ends.
4. The kit of claim 3 , wherein said adaptor overhangs are different and are selected from the group consisting of a, t, c, and g.
5. The kit of claim 1 , wherein said plurality of double stranded nucleic acid adaptors comprises an adaptor overhang comprising c or g.
6. The kit of claim 1 , wherein said adaptor overhangs have double nucleotide overhangs at the 3′ or 5′ ends.
7. The kit of claim 1 , wherein two or more adaptors comprise adaptor overhangs that are different and are selected from the group consisting of aa, ag, at, ac, ga, gg, gc, gt, tt, tc, ta, tg, cc, ct, ca, and cg.
8. The kit of claim 1 , wherein said plurality of adaptors comprises 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, 12 or more, 13 or more, 14 or more, 15 or more or 16 or more distinct overhangs.
9. The kit of claim 1 , wherein said plurality of double stranded nucleic acid adaptors comprises:
a) a first double stranded nucleic acid adaptor comprising an adaptor overhang of 1 or 2 nucleotides;
b) a second double stranded nucleic acid adaptor comprising an adaptor overhang of 1 or 2 nucleotides; and
c) a third double stranded nucleic acid adaptor comprising an adaptor overhang of 1 or 2 nucleotides,
wherein:
i) said adaptor overhang of said first double stranded nucleic acid adaptor is different than said adaptor overhang of said second double stranded nucleic acid adaptor; and
ii) each of said adaptor overhangs of said first and second double stranded nucleic acid adaptors is different than said adaptor overhang of said third double stranded nucleic acid adaptor.
10. The kit of claim 1 , wherein two or more double stranded nucleic acid inserts comprise distinct nucleic acid sequences.
11. The kit of claim 1 , wherein said single stranded nucleotide overhangs of said one or more double stranded nucleic acid inserts comprise a plurality of distinct sequences.
12. The kit of claim 1 , wherein said single stranded nucleotide overhangs of said one or more double stranded nucleic acid inserts comprise overhangs of 1 or 2 nucleotides.
13. The kit of claim 1 , further comprising at least one reagent for analyzing said one or more double stranded nucleic acid inserts.
14. The kit of claim 13 , wherein said reagent is a reagent for nucleic acid sequencing selected from the group consisting of pyrosequencing, sequencing-by-ligation, sequence-by-synthesis (SBS), nanopore sequencing, and oligonucleotide extension sequencing.
15. The kit of claim 1 , wherein each adaptor of said plurality of double stranded nucleic acid adaptors comprises a detection label.