GSM1486403: Kidney DNase-seq Rep 1; 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 kidney dissected and stored in liquid nitrogen.  0.5-1.0g of frozen tissue (2 kidneys) 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
GSM1486403