title: GSM5048323: Aged_Phf6-WT_3; Mus musculus; RNA-Seq
library_construction_protocol: Fresh LT-HSCs (Lin- Sca-1+ CD117+ CD135- CD150+ CD48-) were FACS-sorted from lineage depleted (mouse lineage depletion kit; Miltenyi Biotec) bone marrow harvested from Phf6-WT and Phf6-KO mice at 16-weeks and 24-months of age. Cells were sorted directly into RLT buffer and total RNA was extracted using the RNAeasy mini kit (Qiagen). Total RNA concentration was determined using the Qubit RNA HS assay and the RNA Integrity Number (RIN) was measured using the Bioanalyzer system (Agilent) for each total RNA isolate. Only samples with a RIN greater than 8 were used to construct RNA-seq libraries.  RNA-seq libraries were prepared using the SMART-Seq v4 Ultra Low Input RNA Kit for Sequencing (Clontech Laboratories; cat. #634888) and sequencing-ready libraries were generated using the Low Input Library Prep Kit v2 (Clontech Laboratories; cat. #634899). Library concentrations were determined using the Qubit dsDNA HS Assay Kit (Invitrogen; cat. #Q32854)