GSM5121434: LB10_HMO; Mus musculus; RNA-Seq
Total RNA was isolated from homogenized tissue and cells and purified using E.Z.N.A. Hp Total RNA kit (Omega Tek) or RNeasy Mini kit (Qiagen) according to the manufacturer's instructions. The quality and quantity of the total RNA was monitored and measured with NanoDrop (NanoDrop Technologies, Inc. Wilmington, DE). For quantitative PCR analysis, 5-10 ng of cDNA was used for real-time PCR with gene-specific primers (Supplemental Table 2) and TBP as a house keeping gene on a BioRad CFX96 Real-time PCR system (Bio Rad). Transcriptomes of quiescent BMDM (PBS) were compared to BMDM incubated with either 3'SL or LPS alone and LPS and 3'SL together. RNA was extracted from cells and stranded RNA-Seq libraries were prepared from polyA enriched mRNA using the TruSeq Stranded mRNA library prep kit (Illumina). Library construction and sequencing was performed by the University of California San Diego (UCSD) Institute for Genomic Medicine.