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

Introduction

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.

Project Aim

This project aims at bridging the gap between long reads and high throughput by developing a fragment labelling system compatible with high-throughput short-read Illumina sequencing technology.

Labelling System

The labelling system consists of tags that can be incorporated into the target nucleic acid sequences using Tn5-transposase. These tags can be subsequently used to create fragments from the target molecule, allowing the generated fragments to retain information about their original position in the target molecule. This enables the reconstruction of the sequence of the original, individual target molecules from them.

Market Potential

This technology has considerable market potential due to its relatively high ease of market entry. It can be incorporated into existing library preparation techniques for next generation sequencing since many of them readily use Tn5 transposase for fragment generation.

Applications

Additionally, the ability 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. This technology allows for:

1. Complete reconstruction of diploid human genomes by tackling more difficult, highly repetitive genomic segments.
2. Assembly of more complete genomes from clinical samples in the field of metagenomics.
3. Provision of information about isoform- and allelic-specific expression through the reconstruction of full-length transcripts in the field of transcriptomics.