Illumina HiSeq 2500 sequencing; RNAseq of murine primary bone marrow cells transformed by an inducible Hoxa9-ER: time-course
RNAseq of murine primary bone marrow cells transformed by an inducible Hoxa9-ER: time-course
Primary hematopoietic precursor cells from C57/BL6 age 3 to 6 months old female mice were transformed in vitro by an tamoxifen inducible version of Hoxa9 (Hoxa9-ER). Samples were generated in the presence of active Hoxa9 (0h) and in a time series after Hoxa9 was inactivated by tamoxifen withdrawal at 8h, 16h, 24h, 48h, and 72h. nascent RNA isolation, cells were labeled with 4-thio-uridine for 1h before total RNA was extracted by standard methods. Labeled RNA was specifically biotinylated and separated by streptavidin chromatography. Selected RNA was recovered by cleavage of biotion crosslinkers and subjected to standard library construction methods.  Illumina standard RNA library construction kit using Illumina TrueSeq RNA Library Preparation Kit
Experimental Factor: time
0
Experimental Factor: stimulus
tamoxifen