GSM4145676: unstimulated rep2; Mus musculus; ChIP-Seq
ChIP was performed as previously described (Mir et al., Methods Mol Biol 1966: 39-70, 2019) with minor modifications. Briefly, for each ChIP-seq sample  around 1g of frozen muscle tissue (pooled gastrocnemius muscles from 6 different mice) was homogenized, filtered through a 70µm cell strainer, and pellet fixed in 1% Formaldehyde. Nuclei were purified and chromatin sonicated to a 1-0.5 kb size by using Diagenode Bioruptor Plus. Sheared chromatin was then immunoprecipitated with anti-H3S10ph antibodies (Active Motif; Lot No:08308001), and ChIP DNA was then purified and sequenced. Libraries from ChIP DNA were prepared by use of KAPA Hyperprep Kit (Kapa Biosystems, KK8504), Illumina compatible adapters were synthesized by IDT (Integrated DNA Technologies) and used at a final concentration of 68 nM. Adapted ligated libraries were size selected (360-610 bp) in a Pippin Gel station (Sage Science) using 2% dye free gels (Sage Science, CDF2010). Libraries concentration was estimated by Real Time PCR, with KAPA Library Quantification Kit (Kapa Biosystems, KK4873). Quality of the libraries was evaluated by use of the Agilent High Sensitivity DNA Kit (Agilent) in a 2100 Bioanalyzer (Agilent).
GEO Accession
GSM4145676