title: GSM2259990: Adelman_Bl6_mESC_C2cells_2i_MNase-seq_rep2; Mus musculus; MNase-Seq
library_construction_protocol: Nuclei were pelleted at 1000 X G for 5 minutes and 5 × 106 nuclei were resuspended in 400 μl digest buffer (15 mM Tris pH 8, 60 mM KCl, 15 mM NaCl, 1 mM CaCl2, 0.25 M sucrose, 0.5 mM DTT), and incubated with varying concentrations of MNase (Worthington) for 5 minutes. 400 μl stop solution (1% SDS, 0.1 M sodium bicarbonate, 20 mM EDTA) was added and digested nuclei were incubated at 65°C for 90 minutes. 24 μl Tris pH 7.6, 3 μl Proteinase K (Life Technologies), and 3 μl GlycoBlue (Life Technologies) were added and digestions were incubated for 12 hours at 65°C. DNA was extracted and mono-nucleosome fragments were gel-purified from fractions digested to ~60% mono-nucleosome, ~30% di-nucleosome, and ~10% tri-nucleosome- sized fragments. MNase-seq libraries were prepared as described in (Gilchrist et al., 2010)
attributes.tag: GEO Accession
attributes.value: GSM2259990