For MNase digestion, olfactory neuroepithelia were harvested from Mecp2 KO and WT littermate adult mice and resuspended in PIPES buffer (5mM PIPES, 85mM KCl, 0.5% NP-40, pH 8.0) at 4 °C. After disruption with a dounce homogenizer, nuclei were collected by centrifugation. Collected nuclei were washed once, resuspended in the MNase buffer and digested with 0.5 units of MNase (New England Biolabs, Ipswich, MA) per microliter volume for 15 min at 37 °C. MNase digestion was stopped by putting the samples on ice and adding EDTA to a concentration of 10 mM. After digestion with 0.1 µg/µl RNase A (Fermentas, Pittsburgh, PA), DNAs were purified with DNA Purification Magnetic Beads (Life Technologies, Grand Island, NY), and pellets were dissolved in H2O. DNA fragments corresponding to mononucleosomes (about 150 bp) were confirmed by using 2100 Bioanalyzer (Agilent Technologies Santa Clara, CA). MNase-seq libraries were prepared as described (Bioo Scientific, Austin, TX) using 5140-01 NEXTflex DNA Sequencing kit and 514101 NEXTflex DNA Barcodes-6. High throughput sequencing was done with Illumina Hi-Seq 2000. library construction protocol
GEO Accession
GSM1827608