GSM3756105: Female_B6_DNaseI_Repl2_G163_M7; Mus musculus; DNase-Hypersensitivity
Liver was homogenized, nuclei were suspended in a high sucrose buffer, and then ultra centrifuged to obtain purified intact nuclei. 50 million liver nuclei per sample were incubated with 32 U of RQ1 RNase-free DNase I (Promega, cat#:M6101) for 2 min at 37C. From the purified DNA, fragments between 125-400 bp long were isolated using a sucrose gradient and with Agencourt AMPure XP beads (Beckman Coulter, Cat# A63881) size selection. 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
GSM3756105