GSM9039132: HeLa cells, esiDNAJC15_3; Homo sapiens; RNA-Seq
Cells were harvested and washed with ice-cold PBS prior to cell lysis using the lysis buffer provided in the RNA isolation kit (Macherey-Nagel, Cat# 740955.250). The experiment was performed according to the manufacturer's instructions. RNA isolation was quantified using a NanoDrop spectrophotometer (Thermo Fisher, Cat# ND-ONE-W) 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. The library was checked with Qubit and real-time PCR for quantification and bioanalyzer for size distribution detection. Quantified libraries will be pooled and sequenced on Illumina platforms, according to effective library concentration and data amount.