title: GSM5507972: 4t1+Tcells.Vtn.55; Mus musculus; RNA-Seq
library_construction_protocol: Total RNA was harvested from frozen samples using the RNeasy Mini Kit (Qiagen, 74104).  Thawed, pelleted cells were disrupted with the RNeasy Mini Kit RLT lysis buffer containing β-mercaptoethanol and homogenized through a sterile, blunt 20-gauge needle.  On-column DNA digestions were performed using the RNase-Free DNase Set (Qiagen, 79254).  RNA yields were determined on a Qubit 4 Fluorometer using the Qubit RNA HS Assay Kit (ThermoFisher, Q32852).  RNA quality was assessed using an Agilent 2100 Bioanalyzer Instrument and the RNA 6000 Pico kit (Agilent, 5067-15103). DNA libraries were generated from 0.1-1μg total RNA using the TruSeq RNA Sample Prep Kit v2 (Illumina, RS-121-2001) and purified using AMPure XP magnetic beads (Beckman Coulter, A63881).  Libraries were normalized, pooled and sequenced on an Illumina NextSeq 550 System, generating single-end, 75 base pair reads.