Gut microbiome of Tilapia (Oreochromis niloticus) from Atlantic farm in Colombia
DNA was extracted from the seed culture, day 0, following 24-hour incubation of the combined intestinal contents from Nile tiapia and at 1, 2, 3, 5, 17, and 33 days incubation of the CFCEC using the Stool DNA Isolation kit (Norgen, Thorold, ON, Canada) according to the manufacturers instructions. PCR primers 515F/806R with barcodes were used to sequence the highly variable V4 region of the 16S rRNA gene. PCR was carried out with the HotStarTaq Plus Master Mix kit (Qiagen, Germantown, MD, USA) with the following conditions: initial denaturation at 94C for 3 min, 28 cycles at 94C for 30 sec, at 53C for 40 sec, and 72C for 60 secs, and final extension at 72C for 5 min. Amplicons were confirmed using 2% agarose gel electrophoresis. The PCR products were sequenced at Molecular Research DNA Lab (Shallowater, TX, USA) using 300 to 570 bp paired-end sequencing with an Illumina Miseq following the manufacturers guidelines.