GSM4156412: Stimulated_24h_3; Mus musculus; RNA-Seq
Cells were harvested and RNA extracted using Rneasy mini kit from Qiagen according to manufacturer's instructions. RNA sequencing libraries were prepared using the TruSeq stranded mRNA library preparation kit in accordance to the manufacturer instructions. Briefly 500ng of total RNA was used for purification and fragmentation of mRNA. Purified mRNA underwent first and second strand cDNA synthesis. cDNA was then adenylated, ligated to Illumina sequencing adapters and amplified by PCR using 10 cycles. Final libraries were evaluated using fluorescent based assays including PicoGreen (Life Technologies) or Qubit Fluorometer (Invitrogen) and Fragment Analyzer (Advanced Analytics) or BioAnalyzer (Agilent 2100) and were sequenced on an Illumina HiSeq2500 sequencer (v4 chemistry) using 2*125bp cycles.