GSM3109360: BMDM_veh_r1; Mus musculus; RNA-Seq
For bulk RNA-seq of sort-purified cell populations, libraries were prepared using the SMARTer Stranded RNA-seq kit (Clontech) and TruSeq RNA library prep kit (Illumina). For bulk RNA-seq of adipose tissue, total RNA was isolated from snap-frozen tissue samples. Tissues were mechanically homogenized in 500μL TRIzol (Life Technologies) in a TissueLyser (Qiagen) for 4 min, at a frequency of 22s−1. After homogenization, samples were centrifuged at 4°C and homogenates transferred to a fresh tube for RNA extraction with chloroform. Aqueous chloroform fractions were mixed with equal volumes of 70% ethanol/30% water and RNA further purified with RNeasy Mini spin columns (Qiagen) and oncolumn DNase-digestion (Qiagen). One microgram of purified DNase-treated total RNA from biological replicates was processed with a Ribo-Zero Magnetic rRNA removal kit (Epicentre, MRZH11124). Bulk adipose RNA libraries were prepared using a TruSeq Stranded Total RNA Library Prep Gold (Illumina, 200020599) and standard Illumina protocol.
GEO Accession
GSM3109360