mRNA-Seq of mus musculus: adult male spleen
A total amount of 1 g RNA per sample was used as input material for the RNA sample preparations. mRNA from eukaryotic organisms was enriched using oligo(dT) beads from NEBNext Poly(A) mRNA Magnetic Isolation Module (cat# E7490L - NEB, USA). Subsequently, sequencing libraries were generated using NEBNext Ultra II Directional RNA Library Prep Kit for Illumina  (cat# E7770L - NEB, USA) following manufacturers recommendations.The clustering of the index-coded samples was performed on a cBot Cluster Generation System (cat# SY401-2015, Illumina) using TruSeq PE Cluster Kit v3-cBot-HS (cat# PE-401-3001, Illumia) according to the manufacturers instructions. After cluster generation, the libraries were sequenced on an NovaSeq 6000 Illumina platform using NovaSeq 6000 S2 Reagent Kit v1.5 cat. 20028314 -(300 cycles) and 150 bp paired-end reads were generated (minimum 12Gb and 40M).