title: GSM3523235: ChIP-seq_H3K27me3_EProg; Homo sapiens; ChIP-Seq
library_construction_protocol: Chromatin from E-Prog and E-Prec was prepared after cross-linking for 10' at RT with 1% formaldehyde-containing medium. Nuclear extracts were sonicated using the Bioruptor Pico Sonication System (Diagenode) to obtain DNA fragments around 150-200 bp in length. For histone modifications, chromatin obtained from 107 cells was immunoprecipitated overnight with 10 µg of antibody. For transcription factors, chromatin obtained from 3x107 cells was immunoprecipitated overnight with 30 µg of antibody.  ChIP assay was performed as previously described (Romano, Sci Reports, 2016; Cui et al., 2009). ChIP-seq libraries for E-Prog and E-Prec were prepared from 1 ng of immunoprecipitated DNA (IP) and 5 ng of control DNA (Input: non-immunoprecipitated chromatin fragments) following the Diagenode Microplex Library preparation kit. Libraries were checked by capillary electrophoresis by Tape Station Agilent with the High sensitivity D1000 assay and quantified by Real Time q-PCR using the kit from KAPA Biosystems (Roche). Each library was sequenced in one MiSeq Illumina RUN and 50 bp single-end reads were generated.
attributes.tag: GEO Accession
attributes.value: GSM3523235