GSM9320915: Bleo_2W_WholeLung_1 (biological replicate 1); Mus musculus; RNA-Seq
Viable lung cells were enzymatically dissociated into single-cell suspensions, treated with the 10x Genomics proprietary crosslinking reagent to preserve RNA integrity, and stored at –80 °C until all experimental groups were collected. Single-cell RNA-seq libraries were prepared using the 10x Genomics Fixed RNA Profiling (scRNA-seq Flex) chemistry following the manufacturer's protocol. Target transcript capture and sample barcoding were performed using the Chromium Mouse Transcriptome Probe Set v1.0.1 with an additional custom probe targeting EGFP. Barcoded cells were encapsulated into Gel Bead-in-Emulsions (GEMs) using the Chromium Chip Q. Inside each GEM, probe pairs were ligated, and unique molecular identifiers (UMIs), GEM barcodes, and partial Read 1 sequences were incorporated through polymerase extension. Libraries were PCR-amplified and indexed to generate sequencing-ready libraries containing Illumina P5/P7 adapters. Library quality was assessed by Agilent Bioanalyzer and Qubit fluorometer prior to sequencing.