GSM4976660: WT-3_MPP; Mus musculus; RNA-Seq
Murine HSCs and MPPs were FACS sorted from mice under homeostatic conditions. The total RNA concentration from sorted mouse DCs was determined with the NanopDrop 1000 spectrophotometer (NanoDrop, Wilmington, DE) and RNA quality assessed with an Agilent Bioanalyzer 2100 (Agilent, Santa Clara, CA). There were three independent DC samples for each treatment group. The TruSeq RNA Sample Preparation Kit V2 (Illumina, San Diego, CA) was used for next generation sequencing library construction per manufacturer's protocols. Briefly, mRNA was purified from 100ng total RNA with oligo-dT magnetic beads and fragmented. First-strand cDNA synthesis was performed with random hexamer priming followed by second-strand cDNA synthesis. End repair and 3` adenylation was then performed on the double stranded cDNA. Illumina adaptors were ligated to both ends of the cDNA, purified by gel electrophoresis and amplified with PCR primers specific to the adaptor sequences to generate amplicons of approximately 200-500bp in size.