GSM9588998: 20% O2 wo Doxy replicate 1; Homo sapiens; RNA-Seq
Total RNAs were prepared from FDX2-iKO ES2 cells cultured 6 days with or without Doxy at 20% O2 using Maxwell RSC simplyRNA Kits (Promega). Alternatively, cells were cultured for an additional 3 days at 3% O2 after a 4-days of preculture at 20% O2. mRNA 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. 5'-ends of each library were phosphorylated and cyclized. Subsequently, loop amplification was performed to generate DNA nanoballs.