GSM7927212: NCI-H1975 cells, sgDAMP, input_mRNA, rep2; Homo sapiens; RNA-Seq
For mRNA-seq, total RNA was isolated using TRIzol (Invitrogen) and quantified by spectrophotometry. For ribo-seq, samples were lysed and subjected to nuclease digestion using the TruSeq Ribo Profile Library Prep Kit (Illumina) according to the manufacturer's instructions. Lysates were then purified using MicroSpin S-400 HR spin columns (Thermo Fisher). rRNA was then depleted using a Ribo-Zero Plus rRNA Depletion Kit (Illumina). For mRNA-seq, extracted mRNA was enriched with oligo(dT) beads, subjected to fragmentation, and reverse transcribed into cDNA with random primers. cDNA samples were then purified, subjected to end-repair and oligo(A) added and ligated to Illumina adaptors according to the manufacturer's instructions; For Ribo-seq, rRNA-depleted samples were subjected to purification by PAGE, fragmentation, end repair and 3' adapter ligation, reverse transcription, and cDNA circularization following a protocol described by Illumina.