GSM5866502: 4xLPS-1-NO; Mus musculus; RNA-Seq
Total RNA was isolated from BMDMs using the RNeasy Mini Kit (Qiagen). The mRNA was then purified from 500 ng of total RNA using the Dynabeads mRNA DIRECT Micro Kit (Thermo Fisher Scientific). Whole-transcriptome libraries were prepared using the Ion Total RNA-seq Kit v2. The yield and size distribution of the amplified libraries were assessed with the Agilent 2100 Bioanalyzer using the DNA 1000 Kit. Sequencing was performed on an Ion Proton (Thermo Fisher Scientific). Analysis was performed using the Torrent Suite software v5.12 (Thermo Fisher). The whole Transcriptome Analysis Plugin aligns reads on mouse reference genome (mm10) using Tophat2 then unmapped reads are aligned using Bowtie2 and merged together. FPKM are calculate using Cufflinks.