16S rRNA gene amplicon sequencing of global harbor water and wastewater
400 mL of surface water was filtered on a 0.22 micrometer filter, which was then folded into a microcentrifuge tube and stored frozen at -80C until DNA extraction. From DNA extracts, amplicons were generated using PCR primers targeting the v4v5 region of the 16S rRNA gene. Triplicate PCR reactions were pooled, barcoded, and prepared into sequencing libraries for 250bp paired end sequencing on an Illumina MiSeq instrument.