GSM8567829: eAT_5X_6; Mus musculus; RNA-Seq
Total RNA was extracted with TRIzol reagent (Invitrogen) after tissue lysis with stainless steel beads of 5 mm (Qiagen) and purified using RNeasy Mini kit (Qiagen). To construct the libraries, 537 ng of high quality total RNA sample (RIN >7) was processed using Stranded mRNA Prep kit (Illumina) according to manufacturer instructions. Briefly, after purification of poly-A containing mRNA molecules, mRNA molecules are fragmented and reverse- transcribed using random primers. Replacement of dTTP by dUTP during the second strand synthesis will permit to achieve the strand specificity. Addition of a single A base to the cDNA is followed by ligation of Illumina adapters. Libraries were quantified by Q bit and profiles were assessed using the DNA High Sensitivity LabChip kit on an Agilent Bioanalyzer. Libraries were sequenced on an Illumina NovaSeq instrument using 51 base-lengths read in a paired-end mode.