GSM2752308: Male_Rad21chip_Repl2_G133_M6; Mus musculus; ChIP-Seq
Nuclei were isolated by ultracentrifugation in a high sucrose buffer then crosslinked with 0.8% formaldehyde for 9 minutes. Samples were sonicated with a Bioruptor Twin (Diagenode, UCD-400) until the majority of DNA fragments were between 100-400bp (60-70 cycles total, 30 sec ON 30 sec OFF). Chromatin was quantified and 70 ug of chromatinized DNA was incubated with the indicated ChIP antibody. Samples were prepared for sequencing with NEBNext Ultra II DNA Library Prep Kit for Illumina according to the manufacturer's directions for low input samples (New England Biolabs, #E7645). All samples were subjected to double-sided SPRI size selection prior to PCR amplification (Agencourt AMPure XP; Beckman Coulter: A63882). Samples were barcoded using New England Biolabs NEBNext Multiplex Oligos for Illumina (#E7335) with 8 rounds of PCR per the manufacturer's instructions.
GEO Accession
GSM2752308