title: GSM9386813: Fnfs-derived M1 macrophages untreated, Day 3, Biological Replicate 2; Homo sapiens; RNA-Seq
library_construction_protocol: Cells were released from hydrogels (each replicate had 10 hydrogels) using dissolution buffer (55 mM sodium citrate, 50 mM EDTA, 90 mM NaCl, pH 6.9) and homogenized in TRIzol.RNA was isolated using the miRNeasy Micro Kit. Total RNA was extracted from TRIzol lysates and assessed for integrity (RIN ≥ 9.0). Poly(A)+ RNA was enriched using oligo-dT magnetic beads (NEBNext Poly(A) Magnetic Isolation Module). Strand-specific libraries were prepared using the NEBNext Ultra II Directional RNA Library Prep Kit (New England Biolabs), following the manufacturer's protocol. Libraries were quantified using KAPA qPCR and quality-checked on BioAnalyzer or TapeStation (RIN cutoff ≥ 7.0). Libraries were multiplexed and sequenced on Illumina NextSeq 6000 (paired-end 101 bp reads, 101-8-8-101 structure) or NovaSeq 6000 (~25 million paired-end reads per sample).