GSM8751454: J774_wt_ctrl_LPS_rep2; Mus musculus; RNA-Seq
RNA was isolated using the ReliPrep RNA miniprep system (Promega) following the manufacturers instructions. In brief, cells were lysed in BL+TG buffer and lysates stored at -70 °C. For isolation of RNA, lysates were mixed with 2-propanol, bound to a silica membrane washed and DNA was digested by DNAseI. After additional washes RNA was eluted in nuclease-free water. RNA concentration was measured using a Nanophotometer (Implen) and 100-1000 ng of RNA were reverse transcribed using the  Reverse Transcription System with random hexamers (Promega) following the manufacturers instructions. Thereby a control without reverse transcription was included to assess potential DNA contaminations during qPCR. The RNA quality was determined on an Agilent 2100 Bioanalyzer with the RNA 6000 Nano kit, following the manufacturer's instructions. Library preparation and rRNA depletion were conducted using the TruSeq unstranded mRNA Library Prep kit (Illumina) starting with 500 ng of RNA for each biological triplicate. The samples were sequenced on the Illumina NovaSeq 6000 or NovaSeq X PLUS.