Illumina Genome Analyzer II sequencing; DHS NS5 Cells
DHS NS5 Cells
NS5 cells were cultured as previously described (Conti et al. 2005), using laminin as a media supplement (20ug/mL), and plated at a density of 28,000/cm2 NS5 cells were infected with pBABE-IRES-GFP-puro (NS5_control) or pBABE-Ascl1-ERT2-IRES-GFP-puro (NS5_Ascl1), followed by selection in complete medium containing 1ug/mL puromycin (Calbiochem), the latter sample collected 24 hours after induction with 4-Hydroxytamoxifen at a final concentration of 50nM, while reducing EGF concentration to 5ng/mL . DNase-seq samples from NS5 cells were prepared as described by Song et al (Genome Res, 2011). Libraries were generated as in (Boyle et al., 2008; Song and Crawford, 2010) with a slight modification made to the linkers to increase ligation efficiency (Song et al., 2011, Genome Res).
Experimental Factor: genotype
pBABE-Ascl1-ERT2-IRES-GFP-puro