Next-generation sequencing libraries
Provided herein is technology relating to next-generation sequencing and particularly, but not exclusively, to methods and compositions for preparing a next-generation sequencing library comprising short overlapping DNA fragments and using the library to sequence one or more target nucleic acids.
1. A system for sequencing a target nucleic acid, the system comprising:
a) a nucleic acid ladder fragment library wherein said nucleic acid ladder fragment library comprises a plurality of nucleic acid fragments terminated by a 3′-O-propargyl nucleotide; and
b) a sequencing apparatus.
2. The system of claim 1 further comprising a copper-based click chemistry catalyst reagent.
3. The system of claim 1 further comprising an adaptor oligonucleotide.
4. The system of claim 1 further comprising an adaptor oligonucleotide, wherein the 3′-O-propargyl nucleotide comprises a first reactive group and the adaptor oligonucleotide comprises a second reactive group that is capable of being linked to the first reactive group by click chemistry.
5. The system of claim 1 wherein said nucleic acid ladder fragment ladder library comprises a plurality of nucleic acids having 3′ ends that differ by less than 20 nucleotides.
6. The system of claim 1 further comprising software for assembling short overlapping nucleotide sequences into a consensus sequence.
7. The system of claim 6 wherein:
i) each short overlapping nucleotide sequence comprises less than 100 bases;
ii) the short overlapping nucleotide sequences are tiled over a target nucleic acid comprising at least 100 bases; and
iii) the short overlapping nucleotide sequences are offset from one another by 1-20 bases.