Illumina HiSeq 2500 paired end sequencing; RNA-seq of Smarca5-deficient CD4/CD8 double-positive thymocytes isolated from 6-week mouse thymi and their comparison with wild-type and TP53-null controls.
RNA-seq of Smarca5-deficient CD4/CD8 double-positive thymocytes isolated from 6-week mouse thymi and their comparison with wild-type and TP53-null controls.
DP_H1_p
Double positive CD4+ CD8+ thymocytes from 6-week mice were sorted by FACS. Lineage positive cells (B220+ Gr-1+ Nk1.1+ CD11c+ CD11b+) were excluded from experiment. Total RNA from FACS-sorted DP cells was isolated using TRIzol Reagent  (Invitrogen) following the manufacturers RNA isolation protocol. Strand-specific cDNA libraries were prepared from a minimum of 1.7µg of each DNAse treated (AM1906; Ambion DNA-free Kit) RNA sample using the TruSeq Stranded mRNA LT kit (Illumina). Replicate RNA samples pooled from 6 mice provided sufficient amounts of RNA from S5fl/flhCD2iCre DP cells.