alias: GSM3595865
title: GSM3595865: WT-2; Mus musculus; RNA-Seq
library_construction_protocol: RNA was harvested using Trizol reagent (Invitrogen). Chromatin was prepared from isolated acinar cells or cell lines using standard protocol of ChIP Kit (Active Motif). In brief, cells were fixed by 1% formaldehyde for 10 min and terminated by glycine for 5 min (final concentration = 0.125 M). After 2 times washing by pre-cooling PBS (protease inhibitor containing), cells were scraped and resuspended in SDS-lysis buffer respectively (50 mM Tris-HCl Ph 7.5, 10 mM EDTA, 1% SDS and protease inhibitor), and sonicated to shear crosslinked DNA to about 200-bp. Fifty micrograms of fragmented chromatin was incubated overnight with 2.5 μl of anti-H3K36me3 antibody (Abcam, ab9050) and normal IgG (used as negative control). Complexes were isolated using pre-cleaned protein G agarose (1 hour at 4 ℃). Immunoprecipitated DNA was prepared as a standard illumine library according to Illumina's instructions. libraries were prepared for sequencing using standard Illumina protocols
identifiers.namespace: GEO
identifiers.id: GSM3595865