alias: GSM3530845
title: GSM3530845: HNF6 ChIP Input Foxa1L/L;2L/L AAV Cre 1; Mus musculus; ChIP-Seq
library_construction_protocol: Mice were anesthetized, sacrificed, and subjected to liver perfusion. Whole liver was then digested by collagenase (Roche) and hepatocytes removed and isolated by means of a percoll gradient (18). This yielded a population of relatively pure (>95%) hepatocytes. DNA was isolated from mouse hepatocytes or from snap-frozen mouse tissues and incubated in lysis buffer (25 mM Tris-HCl (pH 8), 2 mM ethylenediaminetetraacetic acid, 0.2% sodium dodecyl sulfate, 200 mM NaCl) supplemented with 300 μg/mL proteinase K (Roche) followed by phenol:chloroform extraction and ethanol precipitation. For ChIP analysis, chromatin was fixed and sheared with the M220 Focused-ultrasonicator™ (Covaris®) using the microTube snap-cap. Ten micrograms of chromatin was then immunoprecipitated with 2 μg of or HNF6 (Santa Cruz, sc-13050) antibodies. Libraries were generated using the NEBNext Ultra™ II DNA Library Prep Kit for Illumina (New England Biolabs).
identifiers.namespace: GEO
identifiers.id: GSM3530845