Full-length 16S rRNA metagenome of ZymoBIOMICS Microbial Community Standard - ONT field run rep b
All procedures described were performed in the Key Largo Marine Research Laboratory (KLMRL; a non-sterile field station) in Key Largo, Florida. DNA was extracted, amplified and purified using the DNeasy PowerSoil Pro kit (Qiagen) according to the manufacturers protocol from three replicates of the ZymoBIOMICS Microbial Community Standard (75 l each) along with a sample of field-collected unfiltered seawater and a negative control of nuclease-free water (1 ml). Library preparation and metabarcoding was performed using the Rapid Barcoding Kit 24 V14 (SQK-RBK114.24) following the ONT protocol Rapid sequencing V14 Amplicon sequencing. The multiplexed library was sequenced on an R10.4.1 flow cell (FLO-MIN114) on the MinION Mk1B monitored by MinKNOW v24.11.10 on a MacBook Pro (Sonoma v14.6.1, Apple M1, 16GB, 8 cores), without real-time basecalling. Sequences were basecalled using Dorado implementing the simplex basecalling model dna_r10.4.1_e8.2_400bps_sup@v4.3.0.