SP4
Surface water samples from five anthropogenically impacted wetland sites (SP1SP5) were filtered through 0.22 m membrane filters to capture microbial biomass. High-molecular-weight environmental DNA was extracted from filters using a commercial DNA extraction kit according to the manufacturers protocol. DNA quality and quantity were assessed by fluorometric quantification and spectrophotometry. SMRTbell libraries were prepared using the SMRTbell Express Template Prep Kit 2.0 (Pacific Biosciences), including DNA damage repair, end repair/A-tailing, adapter ligation, and AMPure PB bead purification. Libraries were quality-checked prior to sequencing. Whole-metagenome shotgun sequencing was performed on the PacBio Sequel II platform (Pacific Biosciences) using SMRT technology. Libraries were sequenced in circular consensus sequencing (CCS) mode to generate high-fidelity (HiFi) reads. CCS reads were generated and quality filtered using SMRT Link software with default parameters.