Illumina HiSeq 2500 paired end sequencing; RNA sequencing comparing different stages of leukemic progression in a mouse model of severe congenital neutropenia (SCN)
RNA sequencing comparing different stages of leukemic progression in a mouse model of severe congenital neutropenia (SCN)
Peripheral blood (PB) was obtained by cheek puncture. Upon sacrifice, BM was isolated from femurs and tibias of the mice. Cells were harvested by flushing the bones and filtered. Subsequently, lineage- c-Kit+ (LK) cells were FACS sorted. LK cells were directly sorted in TRIzol (Thermo Fisher Scientific) and RNA was isolated according to the manufacturer's protocol, with the addition of GenEluteTM−LPA(Sigma-Aldrich). cDNA was generated with the SMARTer Ultra Low Input RNA kit for sequencing (version 4, Clontech).