title: LPS2
design: Total RNA was extracted with TRIzol reagent (Invitrogen), and RNA concentration and purity were determined using a NanoDrop spectrophotometer (Thermo Fisher Scientific). Integrity was assessed with an RNA Nano 6000 Assay Kit on the Agilent Bioanalyzer system. mRNA was purified from total RNA using poly-T oligo-conjugated magnetic beads, then fragmented in First Strand Synthesis Reaction Buffer at elevated temperature. First-strand cDNA was synthesized with random hexamer primers and M-MuLV Reverse Transcriptase (RNase H), followed by second-strand synthesis using DNA Polymerase I and RNase H. After end repair, A-tailing, and adaptor ligation, cDNA fragments of 370420 bp were selected with the AMPure XP system (Beckman Coulter). PCR amplification was performed, and libraries were quality-checked on an Agilent Bioanalyzer 2100 before sequencing on the Illumina NovaSeq platform to generate 150 bp paired-end reads.