From microscale to microbial insights: validating high-throughput microvolume extraction (HiMEx) methods for marine microbial ecology.

ABSTRACT: Extracting and directly amplifying DNA from small-volume, low-biomass samples would enable rapid, ultra-high-throughput analyses, facilitating the study of microbial communities where large-volume sample collection is challenging. This can aid where 'conventional' filtrater-based methods miss capturing smaller microbes, or where microscale variability matters, such as the ocean. Here, we develop and validate physical and chemical-based DNA extractions from microvolumes with universal rRNA gene amplicons and metagenomic sequencing of all domains and viruses, on natural surface seawater and experimentally manipulated marine waters. Compared to 500-mL filter-based extraction, direct PCR of 3 μL of lysate from seawater microvolume extractions ranging from 100-1000 μL consistently captured comp

SUBMITTER: Ghotbi M 

PROVIDER: S-EPMC12721387 | biostudies-literature | 2025 Jan

REPOSITORIES: biostudies-literature