Optimization and evaluation of viral metagenomic amplification and sequencing procedures toward a genome-level resolution of the human fecal DNA virome.

ABSTRACT:

Introduction

Viruses in the human gut have been linked to health and disease. Deciphering the gut virome is dependent on metagenomic sequencing of the virus-like particles (VLPs) purified from the fecal specimens. A major limitation of conventional viral metagenomic sequencing is the low recoverability of viral genomes from the metagenomic dataset.

Objectives

To develop an optimal method for viral amplification and metagenomic sequencing for maximizing the recovery of viral genomes.

Methods

We performed parallel virus enrichment and DNA extraction to generate ∼ 30 viral DNA samples from each of 5 fresh fecal specimens and conducted the experiments including 1) optimizing the cycle number for high-fidelity enzyme-based PCR amplification, 2) evaluating the reproducibili

SUBMITTER: Wang G 

PROVIDER: S-EPMC10248800 | biostudies-literature | 2023 Jun

REPOSITORIES: biostudies-literature