RNA-Seq of glioma:  T98-Glioblastoma
Total RNA was extracted from T98G cells and treated with RQ1 DNase (Promega) to remove DNA. mRNA was captured using VAHTS mRNA capture Beads (Vazyme, N401) or ribosomal RNA was depleted using Ribo-off  rRNA depletion kit (Vazyme, N406-01). The purified RNA was fragmented, and first-strand cDNA synthesis was performed using random primers. Second-strand cDNA synthesis was carried out with dUTP incorporation for strand specificity. The cDNA was end-repaired, A-tailed, and ligated to Illumina adapters. The libraries were size-selected (300-500 bp) and amplified by PCR. Finally, the libraries were quantified using Qubit 4.0 and sequenced on the Illumina Novaseq 6000 platform with PE150 mode.