Single molecule reconstruction for high-throughput, short-read sequencing technologies

Current advanced short-read sequencing technologies and third generation long-read sequencing technologies are still not capable of full molecule reconstruction in a high throughput manner due to their limited read length and throughput respectively.This project aims at bridging the gap between long reads and high throughput by developing a fragment labelling system compatible to high-throughput short-read Illumina sequencing technology.

The labelling system consist of tags that can be incorporated into the target nucleic acid sequences using Tn5-transposase and can be subsequently used to create fragments from the target molecule, allowing the generated fragments to retain information of their original position in the target molecule, thus the reconstruction of the sequence of the original, individual target molecules from them.This technology has considerable market potential due to its relatively high ease of market entry as it can be incorporated in existing library preparation techniques for next generation sequencing since many of them readily use Tn5 transposase for fragment generation.

Additionally, the ability to be able to link the short, high accuracy reads generated by high throughput next generation sequencing technologies can have a large interest in the field of genomics, by allowing the complete reconstruction of diploid human genomes by tackling more difficult, highly repetitive genomic segments, in the field metagenomics, by enabling the assembly of more complete genomes from clinical samples, and also in the field of transcriptomics, by providing information about isoform- and allelic-specific expression through the reconstruction of full length transcripts.