Metagenome of human pleural drainage
DNA quantification was performed using both the Qubit dsDNA HS Assay Kit (Thermo Fisher Scientific, Waltham, MA, USA) and the Agilent High Sensitivity DNA Kit (Agilent, Santa Clara, CA, USA) to ensure accuracy. Subsequently, libraries were prepared using the NEBNext Ultra II DNA Library Prep Kit for Illumina (New England Biolabs, MA, USA) following the manufacturer's protocol. The prepared library underwent quality checks using the Qubit assay and real-time PCR for quantification, as well as the bioanalyzer for size distribution detection. Once the libraries were quantified and deemed suitable, they were pooled together. The pooled libraries were then sequenced using the NovaSeq6000 instrument (Illumina, San Diego, CA, USA) with the NovaSeq 6000 S4 Reagent Kit v1.5 (paired end, 300 cycles).