title: NextSeq 500 paired end sequencing; RNA-seq of H2afj-ko and isogenic WT C57/Bl6-N mouse embryonic fibroblasts in proliferation and after etoposide-induced senescence.
design: RNA-seq of H2afj-ko and isogenic WT C57/Bl6-N mouse embryonic fibroblasts in proliferation and after etoposide-induced senescence.
library_construction_protocol: Cells trypsinized, washed with PBS, and resuspended in 350 µl Machery-Nagel Nucleospin RNA+ Lysis buffer. MEFs were cultivated in DMEM + Gluta-Max (Gibco 31966) + 10%FBS + pen/strep in a 5% carbon dioxyde and 5% oxygen incubator.  MEFs were passed twice before inducing senescence by treatment with 2.5 µM etoposide for 6 days (with media changed after 3 days), followed by 3 days incubation in fresh medium without etoposide Machery-Nagel Nucleospin RNA+ kit protocol. Illumina TruSeq Stranded mRNA kit protocol.