title: GSM6721087: IVT_12hr_2_S44; Homo sapiens; RNA-Seq
library_construction_protocol: After 10 minutes of incubation at room temperature, sample was spun at 20,000xg for 20 minutes at 4°C, then RNA pellet washed with fresh 75% ethanol before drying and resuspending in 40µL water. isolated mRNA was DNase treated using Turbo DNase in reactions consisting of 11µL (~15µg) RNA and 1.5µL Turbo DNase according to manufacturer's protocol. DNase-treated RNA was purified using lithium chloride precipitation as described in mMESSAGE mMACHINE T7 ULTRA Transcription Kit (Invitrogen). 2µg RNA per sample was reverse transcribed into cDNA using SuperScript III First-Strand Synthesis System (Invitrogen) and MPRA RNA-specific primer (Table **, primer #22).  