title: GSM8034821: WT LPS_2; Mus musculus; RNA-Seq
library_construction_protocol: RNA was isolated using the High Pure RNA Isolation Kit (Roche, #11828665001) according to the manufacturer's protocol and stored at ‑80°C. All RNA samples used for further preparation had an RNA integrity number > 9 according to the analysis in a 2100 Bioanalyzer (Agilent) using the RNA 6000 Nano Kit (Agilent, #5067-1513) Libraries were prepared from 500 ng RNA. Poly(A) enrichment was performed on the input total RNA using the NEBNext Poly(A) mRNA Magnetic Isolation Module (New England Biolabs, #E7490) according to the manufacturer's instructions. The cDNA library preparation was conducted with the NEBNext Ultra Directional RNA Library Prep Kit for Illumina (New England Biolabs, #E7420) as recommended by the supplier. In brief, first- and second-strand cDNA synthesis was performed, followed by adapter ligation and PCR amplification of the final library (12 cycles). PCR cleanup was performed using Agencourt AM‑Pure XP beads (Beckmann Coulter, #A63881). Libraries were sequenced for 1x 75 nt on a NextSeq500 (Illumina) sequencer.