AMPLICON of Oreochromis niloticus: intestinal microbiota
The library were created by using illumina high-throughput sequencing of barcoded 16S rRNA genes obtained from the V3-V4 regions of the bacteria 16S ribosomal RNA genes. The libraries were amplified by polymerase chain reaction (PCR) using specific primers. Unique eight-base barcodes were added to each primer to distinguish PCR products. The raw pair-end readings obtained were subjected to quality-control procedures by using the quantitative insights into microbial ecology (QIIME, version 1.17). The qualified reads were clustered to generate operational taxonomic units (OTUs) at the 97% similarity level by using UPARSE (version 7.1). Chimeric sequences were identified and removed using UCHIME. The phylogenetic affiliation of each 16S rRNA gene sequence was analyzed by Ribosomal Database Project (RDP) Classifier against the silva 16S rRNA database using confidence threshold of 70%. Taxonomic richness and diversity estimators were determined using a Mothur software.