GSM9813870: PBS-treated L929 fibroblast, RNA-seq replicate 1; Mus musculus; RNA-Seq
Total RNA was extracted from L929 cells using the TRIzol reagent (Invitrogen) following the manufacturer's instructions. RNA integrity was assessed using an Agilent 2100 Bioanalyzer, and RNA concentration was quantified using a Qubit Fluorometer (Invitrogen). Only RNA samples with RIN > 8.0 were used for library construction. Sequencing libraries were generated using the NEBNext Ultra II RNA Library Prep Kit for Illumina following the manufacturer's instructions. Briefly, polyA-containing mRNA was enriched from total RNA using oligo(dT) beads, followed by fragmentation, first-strand and second-strand cDNA synthesis, end-repair, 3' adenylation, and ligation with Illumina-compatible adaptors. The adaptor-ligated fragments were then enriched by limited-cycle PCR amplification. Library quality and size distribution were assessed using an Agilent 2100 Bioanalyzer, and quantified by qPCR.