GSM8776990: M2 macrophages, Untreated, rep#3; Homo sapiens; RNA-Seq
Both floating and adherent cultures were collected. RNA was extracted using Rneasy mini kit (Quiagen). Messenger RNAwaspurified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers. Then the second strand cDNA was synthesized using dUTP, instead of dTTP. The directional library was ready after end repair, A-tailing, adapter ligation, size selection, amplification, and purification. The library was checked with Qubit and real-time PCR for quantification and bioanalyzer for size distribution detection.