Linked-read sequencing library preparation
Overview
Executive Statement:
Innovative methods for generating sequence-linked DNA fragments for high-quality, low-cost genome sequencing.
Description:
This technology describes a novel strategy to construct a long-read sequencing library such that adjacent fragments are linked with end-terminal duplications. The CRISPR-Cas9 nickase enzyme and a pool of multiple sgRNAs are used to perform non-random fragmentation of targeted long DNA molecules (>300kb) into smaller library-sized fragments (about 20 kbp) in a manner to retain physical linkage information (up to 1000 bp) between adjacent fragments. This sequencing strategy can be readily applied in both PacBio and Oxford Nanopore platforms and is useful for targeted enrichment of hundreds of target genomic regions and where long no-gap contigs need deep sequencing.
Market Applications
- Haplotype-resolved whole genome sequencing
- High-throughput sequencing in genomics
- Structural variant analysis
- Enriched long-read target sequencing
Key Advantages
- Efficient generation of linked-paired end DNA fragments
- Lower costs in assembling complex genomic regions
- Improved throughput and accuracy of the sequencing
Problems Solved
- Challenges in de novo genome assembly of sequence reads
- Problems with mapping sequencing data to the reference sequence
- Limitations in the resolution of haplotypes
- Enrich the target genomic regions
Intellectual Property and Development Status
United States Patent Pending- #US20240035024A1 - Linked-read sequencing library preparation
Canada Patent Pending - #CA3195700A1 - Linked-read sequencing library preparation
European Patent Pending - #EP4229220A1 - Linked-read sequencing library preparation
References
Nature Scientific Reports 2024: A long-read sequencing strategy with overlapping linkers on adjacent fragments (OLAF-Seq) for targeted resequencing and enrichment
Commercialization Opportunities