title: Illumina Genome Analyzer II sequencing; ChIP NS5 Ascl1-ERT2
design: ChIP NS5 Ascl1-ERT2
library_construction_protocol: 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-Ascl1-ERT2-IRES-GFP-puro (NS5_Ascl1), followed by selection in complete medium containing 1ug/mL puromycin (Calbiochem), and collected  30 minutes after induction with 4-Hydroxytamoxifen at a final concentration of 50nM, while reducing EGF concentration to 5ng/ml NS cells were fixed sequentially with di(N-succimidyl) glutarate and 1% formaldehyde in phosphate buffered saline and then lysed, sonicated and immunoprecipitated, as previously described (Castro et al., 2011), with rabbit anti-HA (ab1424, Abcam) DNA libraries were prepared from 10ng of immunoprecipitated DNA according to the standard Illumina ChIP-seq protocol.
attributes.tag: Experimental Factor: immunoprecipitate
attributes.value: rabbit anti-HA