title: GSM1432735: DSA-2; Mus musculus; RNA-Seq
library_construction_protocol: Small intestinal crypts were isolated in PBS containing 3 mM EDTA, 0.5 mM DTT and protease inhibitor (Roche). Adenomas were dissected from the small intestine. RNA was extracted using RNaEasy kit (Qiagene).For ChIP-seq, crypts and adenoma in 1% formaldehyde for 10 Min at room temperature. Chromatin was sheared using the Covaris S2 system following the manufacturer’s instructions. ChIP was performed using a ChIP validated rabbit anti-Rad21 antibody (Ab992, Abcam) and Magna-ChIP A kit (Millipore). Library preparations were performed using the TruSeq RNA Sample Preparation protocol (Illumina, San Diego, CA) and correct size established using the DNA 1000 kit (Agilent technologies, Santa Clara, CA). Libraries were quantified with qPCR, normalised and pooled to 2nM before sequencing with paired-end 50 bp reads using standard protocols on the HiSeq2000 (Illumina, San Diego, CA).