Metagenomics of D. rerio gut microbiome
PCR amplification was performed using fusion primers targeting from V3 to V4 regions of the 16S rRNA gene with the extracted DNA. For bacterial amplification, fusion primers of 341F (5-AATGATACGGCGACCACCGAGATCTACAC-XXXXXXXX-TCGTCGGCAGCGTC-AGATGTGTATAAGAGACAG-CCTACGGGNGGCWGCAG-3; underlined sequence indicates the target region primer) and 805R (5- CAAGCAGAAGACGGCATACGAGAT-XXXXXXXX-GTCTCGTGGGCTCGG-AGATGTGTATAAGAGACAG-GACTACHVGGGTATCTAATCC-3) were used. The fusion primers are constructed in the following order which is P5 (P7) graft binding, i5 (i12) index, Nextera consensus, Sequencing adaptor, and Target region sequence.