GSM9014164: abc_input_1; Mus musculus; RNA-Seq
RNA (before/after TRAP) was isolated using the Trizol/Pure Link RNA Kit (Ambion). SMARTer Ultra Low Input kit (Clontech) was used for RNA conversion (5 ng for tissue lysate, 10 ng for EC-TRAP samples). cDNA samples were sheared using the Covaris E220 Focused Ultra-sonicator which was followed by Quant-iT PicoGreen quantification (Thermo Fisher Scientific, catalog number P11496). A total of 5 ng cDNA per sample was used for library preparation based on the KAPA Hyper Prep method (KAPA Biosystems, catalog number KK8504). Following NEXTflex adapter ligation (BIOO Scientific, catalog number 514104), libraries were size-selected based on a double bead purification prior to amplification.