GSM5465735: 117B; Ovis aries; miRNA-Seq
Samples from spleen were aseptically taken from each animal at necropsy and tissue sections were preserved in RNAlater solution (Ambion, Austin, TX, USA) at −80 ⁰C. Total RNA was isolated from spleen tissue using TRIzol Reagent (Invitrogen, Carlsbad, CA, USA) and PureLink RNA Mini Kit (Invitrogen). 60 mg tissue samples were homogenized in 1 ml of TRIzol Reagent using Precellys®24 homogenizer (Bertin Technologies, Montigny-le-Bretonneux, France) combined with 1.4 and 2.8 mm ceramic beads mix lysing tubes (Bertin Technologies). RNA isolation was performed following manufacturer instructions and RNA was suspended in RNase free water and stored at −80 ⁰C. The TruSeq Small RNA library prep kit (Illumina, San Diego, CA, USA) was used for the construction of sequencing libraries. miRNA libraries were prepared for sequencing using standard Illumina protocols.