GSM2348253: J774_Live1_24h; Mus musculus; RNA-Seq
Macrophages were lysed by adding 4 M GTC solution (4 M guanidine thiocyanate, 0.5 % sodium N–Lauroylsarcosine, 0.1M β–mercaptoethanol, 0.5 % Tween 80), and the lysate homogenized using a QIAshredder column (Qiagen). RNA was extracted by BCP phase partitioning and isopropanol precipitation, cleaned with 75% ethanol and treated with Turbo DNase (Ambion, Life Technologies). DNA contamination was checked by PCR, and the RNA was further cleaned using an RNeasy MinElute Cleanup kit (Qiagen). Approximately 500 ng of RNA from each sample was used to prepare individually barcoded strand-specific RNA-seq libraries. rRNA and mitochondrial RNA was removed using Ribo-Zero Gold Removal Magnetic Kit for human/mouse/rat (Epicentre, now Illumina) following the manufacturer's instructions. mRNA was purified using Agencourt RNAClean XP kit. Libraries were constructed using ScriptSeq™ Complete Kit