GSM6380198: ARF_5; Homo sapiens; RNA-Seq
QIAamp RNA Blood Mini Kit (Catalog no. 52304)  was used to extract RNA from human whole blood samples. Extraction was performed according to manufacturer's instructions A modified NEBNext RNA Ultra protocol was used to prepare the libraries for mRNA sequencing. The poly-A containing mRNA molecules was purified using oligo-dTattached magnetic beads. Following purification, the mRNA was fragmented using divalent cations under elevated temperature. Next, the cDNA was synthesized using Reverse transcriptase and random hexamers in a first strand synthesis reaction. Subsequently, the cDNA was converted to doublestranded cDNA in a DNA polymerase based reaction. These DNA molecules were then subjected to a series of enzymatic steps for end repair, A-Tailing and ligation with NEB adapters. Finally, these adapter ligated molecules were enriched in a limited cycle PCR with indexed primers and purified using AMPure beads to create final cDNA library for sequencing.