GSM8864898: Primary senescence, X-ray irradiation, replicate 1; Homo sapiens; RNA-Seq
Single-cell suspensions were prepared, and approximately 2.0 × 10⁴ cells were processed using Chromium™ Single Cell 3' Reagent Kits v2 (10x Genomics, cat #120267), following the manufacturer's instructions. Single cells were encapsulated into Gel Bead-in-Emulsions (GEMs) using the Chromium Controller and the Single Cell A Chip Kit (cat #1000009) and I7 Multiplex Kit (cat #120262). Cell lysis occurred within the GEMs, and mRNA was captured with barcoded oligo-dT primers attached to gel beads. Reverse transcription was performed within the GEMs to generate barcoded cDNA, which was then purified and amplified by PCR. Amplified cDNA was enzymatically fragmented, size-selected, and subjected to end repair and A-tailing. The end-repaired and A-tailed cDNA was then ligated with sequencing adapters, and sample indices were incorporated using the I7 Multiplex Kit (10x Genomics, cat #120262). Final libraries were purified, quantified using an Agilent TapeStation, and sequenced on an Illumina HiSeq 4000 platform, following the manufacturer's protocols.