title: GSM4483518: non3.mrna: nonsense siRNA 3; Homo sapiens; RNA-Seq
library_construction_protocol: Cells were exposed to cycloheximide (0.1mg/mL) prior to detergent lysis and scraping as per the Illumina TruSeq Ribo Profile kit manual. Total RNA was extracted from 300µL of clarified generated from the ribosome profiling methodology. For each sample, 1µg of high quality (RIN ≥ 8.5) total RNA was subjected to library preparation for mRNA sequencing using the TruSeq Stranded mRNA Kit (Illumina), as per the manufacturer's instructions. Briefly, mRNA was denatured (65°C, 5 min) and ligated to oligo-dT magnetic beads (room temperature, 5 min), after which unbound RNA and non-specifically bound rRNA was discarded. mRNA was then heat fragmented (94°C, 8 min), following which reverse transcription and adapter ligation were performed as per the manufacturer's instructions. Enrichment of cDNA fragments was subsequently conducted via 15 cycles of PCR, after which libraries were normalised, pooled and subjected to 151 cycles of sequencing using the NextSeq 500 system.
attributes.tag: GEO Accession
attributes.value: GSM4483518