GSM7893372: 122-B_S8; Ovis aries; RNA-Seq
Samples of 1 g of spleen from each sheep were taken at necropsy for RNA extraction and preserved in RNAlater solution (Ambion, Austin, TX, USA) at -80 ºC. Total RNA was isolated from spleen tissue using TRIzol reagent (Invitrogen, Carlsbad, CA, USA) and PureLink RNA Mini Kit (Invitrogen). 60 mg spleen tissue were homogenized in 1 mil of TRIzol Reagent using PrecellysR24 homogenizer (Bertin Technologies, Montigny-le-Bretonneux, France) combined with 1.4 and 2.8 mm ceramic beadsmix lying tubes (Bertin Technologies). RNA isolation was performed following manufacturer instructions and RNA was suspended in RNAse free water and stored at -80ºC. RNA quantity and purity were assesed with NanoDrop 1000 Spectrophotometer (Thermo Scientific Inc, Bremen, Germany). RNA integrity wa assessed on an Agilent 2100 Bioanalyzer with Agilent RNA 6000 Nanochips (Agilent Technologies, Santa Clara, CA, USA), which estimates the 28S/18S (ribosomic RNAs) ration and the RNA integrity number (RIN value). The samples presented and average RIN value of 8.32 and a 260/280 ratio >1,7. RNA libraries were prepared for sequencing using Illumina Stranded Total RNA Prep with Ribo-Zero Plus protocol