Real- time quantiative PCR analysis of HSC derived from sleep deprived mice

HSC were derived from mice sleep deprived for 4 hours and compared to mice that slept for 4 hours during the same circadian time. 5 mice were pooled for each sample and bones from femur, tibia and spine were crushed and stained to lineage deplete and enrich for ckit+, sca-1+,Flk-,CD150+, CD34- qpcr miRNA profiling from approximately 50ng total RNA extracted from HSC FACS sorted into Trizol.