Sea lice microbiome samples collected in four different months from an aquaculture facility in Ireland and sequenced in two different platforms (Illumina and ONT)
The full-length 16S rRNA gene (V1V9; primers 27F/1492R) was amplified using LongAmp Hot Start Taq 2 Master Mix in 25 l reactions (95 C 1 min; 25 cycles of 95 C 20 s, 52 C 30 s, 65 C 2 min; final 65 C 5 min). Amplicons were purified with AMPure XP beads (0.4 ratio), eluted in 30 l nuclease-free water, checked on 1.5% TAE agarose gels, and quantified with PicoGreen.Nanopore libraries were prepared with the Native Barcoding Kit 96 V14 following ONT protocol NBA_9170_v114_revP_30Jan2025. End-prepped amplicons were ligated to unique barcodes, and 20 samples plus positive (ZymoBiomics HMW DNA) and negative controls were pooled and quantified. After adapter ligation and bead cleanup, 50 fmol of library were loaded onto a FLO-PRO114M flow cell and sequenced on a PromethION using MinKNOW v6.4.12, targeting ~300,000 reads per sample.
SUB15807509
Lepeophtheirus_salmonis_metagenome_ONT_May_4