GSM9181226: BT549-control1; Homo sapiens; RNA-Seq
Total RNA was extracted from MCF-7 and BT-549 cells using the QIAGEN RNeasy Mini Kit according to the manufacturer's instructions. Briefly, cells were lysed in RLT buffer containing β-mercaptoethanol and homogenized by pipetting. The lysate was then applied to RNeasy spin columns, followed by on-column DNase I treatment (QIAGEN RNase-Free DNase Set) to remove genomic DNA. After a series of wash steps with RW1 and RPE buffers, total RNA was eluted in RNase-free water. RNA quantity and purity were assessed using a NanoDrop spectrophotometer. Messenger RNA was purified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers followed by the second strand cDNA synthesis. The library was ready after end repair, A-tailing, adapter ligation, size selection, amplification, and purification.