RNA profiling in mouse haemopoiesis

RNA profiling of mouse haemopoetic lineages Overall design: RNA profiling of mouse haemopoetic lineages. For the isolation and sequencing of ouse granulocyte nuclei, bone marrow from C57BL/6J was harvested from the femur, tibia and spine using a mortar and pestle in PBS supplemented with 2% (v/v) FCS and mature granulocytes purified by flow cytometry as described previously. Purification was validated by reanalysis by flow cytometry and May-Grünwald Giemsa staining. Nuclear purification was carried out using the PARIS kit (Ambion). RNA was extracted from the nuclear fraction using Trizol before deep sequencing. Deep sequencing was performed by GeneWorks (Adelaide, Australia) on the Illumina GAII according to the manufacturer''s instructions. For more details, see Supplementary Methods at the NSMB website. Additional files and resources can be found at http://matticklab.com/index.php?title=NuclearTinyRNAs