title: RNAseq of Nile tilapia alevin M10: alevin head kidney
design: After five days of the mentioned treatments administration, Control (commercial feed), A12 (commercial feed + L. lactis A12), M4 (commercial feed + P. megaterium M4), M10 (commercial feed + Priestia sp. M10), A12+M4+M10 (commercial feed + L. lactis A12, P. megaterium M4, and Priestia sp.  M10), and M4+M10 (commercial feed + P. megaterium M4, and Priestia sp. M10), five fish from each treatment were euthanized with an overdose of tricaine methanesulfonate (MS-222). External disinfection of the fish was performed with 70% ethanol. For RNAseq sequence analysis, the kidney was removed aseptically and stored in sterile tubes with TRIZOL at -80C for RNAseq analysis.  According to the manufacturers protocol, the total RNA was extracted from the samples with TRIzol reagent (Invitrogen, Carlsbad, CA). Next, total RNA quantification, quality assessment, rRNA depletion, library preparation, and RNA sequencing were carried out by Novogene Co., Ltd. (Sacrament, CA, USA). In summary, total RNA QC was run for sample quantification (Nanodrop and Agilent 2100), sample integrity (Agilent 2100), and sample purity (Agarose gel and Agilent 2100). Then, according to the manufacturers instructions, total RNA was subjected to rRNA-depletion treatment using a Ribo-ZeroTM Magnetic kit (Illumina). An RNA-Seq library was then constructed using a Next Ultra Directional RNA Library Prep Kit (NEB, Ipswich, USA) and sequenced on an Illumina Novaseq Platform PE150.