Metabarcoding of Oncorhynchus mykiss
Libraries were prepared by following Illumina 16S Metagenomic Sequencing Library Preparation protocol in two amplification steps: an initial PCR amplification using locus-specific PCR primers with overhangs 16S-341F 5- TCGTCGGCAGCGTCAGATGTGTATAAGAGACAGCCTACGGGNGGCWGCAG -3 and 16S-805R 5- GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAGGACTACNVGGGTATCTAATCC -3, and a subsequent amplification that integrates relevant flow-cell binding domains and unique indices (NexteraXT Index Kit, FC1311001/FC1311002). PNA clamping was applied during the first amplification step to block host mitochondrial 16S sequences amplification following the manufacturers protocol (PNA Bio Inc, Newbury Park, CA). Libraries were sequenced on a NovaSeq 6000 instrument (Illumina, San Diego, CA) in paired-end 250-bp mode.
SUB14818982
100997_ID2730_9-A05_S714_L002