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Advancing biosynthetic pathway discovery through short-read-directed long-read sequencing.

Created on 21 Sep 2026

Authors

Marco Gabrielli, Agathe Bredel, Lucas Paoli, Serina L Robinson

Published in

Natural product reports. Sep 21, 2026. Epub Sep 21, 2026.

Abstract

Time span of literature: 2020-todayMetagenomic methods have rapidly advanced, enabling the identification of biosynthetic pathways directly from complex microbiome data. Short-read sequencing, while accurate and cost-effective, often generates fragmented assemblies that can lead to incomplete biosynthetic gene cluster (BGC) recovery. Although long-read sequencing offers a solution to the fragmentation problems, technical requirements and higher costs have limited its scalability. Here, we examine BGC fragmentation in short-read sequencing data across large databases of metagenome-assembled genomes (MAGs) and estimate the targeted genome contiguity required to recover 'complete' biosynthetic gene clusters. We argue that the increasing availability of MAGs recovered from short-read metagenomes with recent advancements in ultra-low input DNA amplification for high-fidelity PacBio sequencing-now requiring as little as nanograms of DNA-can be used sequentially to boost biosynthetic pathway discovery. We demonstrate how natural products researchers can benefit from using short-read MAG comparisons to guide targeted long-read re-sequencing efforts with low amounts of input DNA and/or limited financial resources. Our analysis provides strategic recommendations for the broader scientific community on how to best leverage the strengths of short- and long-read sequencing data to efficiently allocate resources and accelerate natural product discovery.

PMID:
42765534
Bibliographic data and abstract were imported from PubMed on 21 Sep 2026.

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