title: Illumina HiSeq 2000 sequencing; Histone ChIP NS5 Ascl1-ERT2
design: Histone 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 24 hours after induction with 4-Hydroxytamoxifen at a final concentration of 50nM 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 antibodies anti-H3K27Ac and anti-H3K4me1 (ab4729 and ab8895 from 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: anti-H3k4-1Me
attributes.tag: Experimental Factor: phenotype
attributes.value: Ascl1-ERT2 expressed and Tamoxifen-activated
attributes.tag: LIBRARY_STRAND
attributes.value: not applicable