GSM5509592: SLAM_seq_DPY30mAID_Kdm5dKO_48h_rep1; Mus musculus; RNA-Seq
RNA (1 μg) was treated with 10 mM iodoacetamide in a 50-μl reaction volume at 50 °C for 15 min with 50 mM NaPO4 (pH 8.0), and 50% (v/v) DMSO followed by addition of 1 μl of 1 M DTT to stop the reaction. RNA was precipitated at -80 °C for 60 min with 1 μl of GlycoBlue (Thermofisher Scientific, AM9515), 5 μl of 3 M NaOAc (pH 5.2), and 180 μl of 100% ethanol. RNA was pelleted at 4 °C (12,000g, 30 min), washed with 1 mL of 80% ethanol, and centrifuged at 4 °C (12,000g, 30 min). The RNA pellet was dried at room temperature for 10 min and resuspended in 20 μl of nuclease-free H2O. RNA yield and quality were assessed using the 2200 TapeStation (Agilent). Libraries were constructed using QuantSeq 3′-mRNA Seq Library Prep Kit FWD for Illumina (Lexogen, SKU: 015.96).