H3K36me3_ICeChIP-seq_mESC-E14
Mus musculus
source_name
Embryonic stem cells
cell line
E14
chip antibody
H3K36me3 (Abcam, 9050, lot GR165936-1
growth protocol
Mouse Embryonic Stem Cells (mESC) E14 cell line (129/Ola background) (Hooper et al., 1987) were grown at 37°C, 5% CO2, 95% humidity in ES media high glucose DMEM (Invitrogen) supplemented with 15% (v/v) FBS (Gibco), 2mM L-glutamine (Gibco), 1% (v/v) non-essential amino acids (Gibco), 1X Penicillin/Streptomycin (Gibco), 0.1 mM 2-mercaptoethanol (Gibco), 1000U/mL LIF (ESG1107, Millipore), 3µM CHIR99021 (04-0004, Stemgent), 1µM PD0325901 (04-0006, Stemgent), sterile filtered 0.1µm membrane, stored in 4°C for up to 1 week. E14 cells were cultured on dishes coated with 0.1% gelatin (Sigma) without feeder cells, and passaged daily in 1:3 ratio (media was changed 3 hours prior passage). Cells were harvested at 80-90% confluence, with media changed at least 3 hours prior.
amplification cycles
9
map quality cut-off
20
mononucleosome cut-off
200
reference genome
mm9