Sequence tag directed subassembly of short sequencing reads into long sequencing reads

摘要

The invention provides methods for preparing DNA sequencing libraries by assembling short read sequencing data into longer contiguous sequences for genome assembly, full length cDNA sequencing, metagenomics, and the analysis of repetitive sequences of assembled genomes.

主权项

1. A method for generating sequence assemblies from short sequencing reads, comprising:
a) introducing a fragmentation breakpoint in at least one member of an input library to produce a plurality of linear DNA fragments, wherein a first fragment end and a second fragment end are proximal to the fragmentation breakpoint, b) attaching common nucleic acid adaptors to the first fragment end and the second fragment end proximal to the fragmentation breakpoint, wherein the common adaptors attached to the first fragment end and the second fragment end comprise the same unique sequence tag, c) optionally amplifying the plurality of linear DNA fragments to produce a sequencing library comprising a plurality of amplified DNA fragments, wherein at least one of the plurality of amplified DNA fragments comprises:
i) sequence complementary to at least the unique sequence tag of an adaptor, andii) sequence complementary to at least a portion of a member of the input library, d) sequencing at least a portion of the DNA fragments, wherein the presence of a unique adaptor sequence tag in a plurality of fragment sequences thereby associates the fragment sequences having ends that were proximal to the same fragmentation breakpoint, and e) assembling the plurality of breakpoint-associated fragment sequences to generate longer subassembly sequences of the input library.