title: GSM6721939: H3K27ac ChIP-seq denervation gastrocnemius 1 input; Mus musculus; ChIP-Seq
library_construction_protocol: The ChIP seq was prepared using SimpleChIP® Chromatin Immunoprecipitation kit (Magnetic Beads, Cell signaling technology, Cat#9003). Briefly, whole gastrocnemius muscles (500mg) from mice with or without de sciatic denervation were homogenized with dunce grander to isolate myonuclei which were then cross-linked with 1% formaldehyde for 10 min. The myonuclei were treated with 0.5 µl Micrococcal Nuclease for 10 min followed by sonication for 15 min (1/8-inch probe, 30-sec pulses X15 with setting level at 3). After centrifuge, 50ug linked DNA was used for chromatin Immunoprecipitation using anti H3K27Ac antibody (#8173). The pulled down DNA and input DNA were submitted to LC Sciences LLC (Houston, TX. lcsciences.com) for library preparation and the yield library was sequenced with Illumina platform.