GSM7966868: Spleen HSPC, Young, MPL-/-; Mus musculus; RNA-Seq
Marrow and spleen Lin-cKit+ hematopoietic stem/progenitor cells (HSPCs) were isolated by magnetic bead isolation (with ~95% purity) (Miltenyi Biotec) from 3-4mo old wild-type (n=2), TPO-/- (n=3), and MPL-/- (n=3) mice. Total RNA was extracted using the RNeasy mini kit (Qiagen, Hilden, Germany). RNA integrity and quantitation were assessed using the RNA Nano 6000 Assay Kit of the Bioanalyzer 2100 system (Agilent Technologies, CA, USA). For each sample, 400ng of RNA was used to generate sequencing libraries using NEBNextÒ Ultra™ RNA Library Prep Kit for Illumina® (New England BioLabs, MA, USA) following manufacturer's recommendations. The clustering of the index-coded samples was performed on a cBot Cluster Generation System using PE Cluster Kit cBot-HS (Illumina) according to the manufacturer's instructions. After cluster generation, the library preparations were sequenced on an Illumina platform.