RNA-Seq of human glioma cell line U87: ASPHD1 overexpression group3_1
Total RNA integrity was assessed with the RNA Nano 6000 Assay Kit using an Agilent 2100 Bioanalyzer. mRNA was purified with poly-T oligo magnetic beads, fragmented, and reverse-transcribed to first-strand cDNA with random hexamers. Second-strand synthesis used DNA Polymerase I and RNase H. After end repair, A-tailing, adapter ligation, and fragment size selection (370420 bp) with AMPure XP, PCR amplification was performed. The final libraries were validated on an Agilent 2100 Bioanalyzer and sequenced on an Illumina NovaSeq platform (150 bp paired-end).