GSM8285663: U2OS cells G3C2 b; Homo sapiens; RNA-Seq
Total RNA was isolated from log growing U2OS cells (65-75% confluency) using RNeasy mini kit (Qiagen #74104) following manufacturer's guidelines. Library preparation was done by Novogene - mRNA was enriched from total mRNA using oligo(dT) beads. The mRNA was then fragmented randomly in fragmentation buffer, followed by cDNA synthesis using random hexamers and reverse transcriptase. After first-strand synthesis, a custom second-strand synthesis buffer (Illumina) was added with dNTPs, RNase H and Escherichia coli polymerase I to generate the second strand by nick translation. The final cDNA library was ready after a round of purification, terminal repair, A-tailing, ligation of sequencing adapters, size selection and PCR enrichment.