GSM1486404: Liver DNase-seq Rep1; Mus musculus; DNase-Hypersensitivity
25-50 million nuclei were digested with a range of recombinant DNase I enzyme (Roche) concentrations (between 1.2 and 12U) for 15 minutes at 37°C in 120uL 1X DNase buffer. Digestions were checked by pulse field gel electrophoresis and material was pooled from 3 different DNase concentrations (extent of digestion matched between samples) in equimolar amounts following blunt-ending reactions. This was followed by ligation to adapters, MmeI digestion, streptavidin bead-based enrichment, 14 cycles of PCR amplification, and separation from linker-only bands on a gel. C57BL/6 adult female mice were sacked and liver dissected and stored in liquid nitrogen.  0.5-1.0g of frozen tissue (1-2 livers) was ground to fine powder in liquid nitrogen, resuspended in ice cold 20mL nuclei isolation buffer (NIB), filtered through Miracloth, spun down, resuspended in NIB with 0.5% Triton X-100, and centrifuged again to obtain nuclei pellet.
GEO Accession
GSM1486404