title: GSM1301669: ZT8 CLOCK ChIP Seq-1; Mus musculus; ChIP-Seq
library_construction_protocol: livers from mice were immediately homogenized and cross-linked in PBS buffer including 1% formaldehyde (5 ml per liver), and the homogenate was incubated for 15 min at 25 °C. Cross-linking reactions were stopped by the addition of 21 ml of ice-cold nuclei buffer (2.2 M sucrose, 125 mM glycine, 10 mM HEPES pH 7.6, 15 mM KCl, 2 mM EDTA, 0.15 mM spermine, 0.5 mM spermidine, 0.5 mM DTT, 0.5 mM PMSF and 10% glycerol). The homogenate was layered on top of 10 ml of cold nuclei buffer and centrifuged for 90 min at 24,000 rpm (100,000g) at 4 °C in a Beckmann SW27 rotor. The nuclei were washed with wash buffer (20 mM Tris, pH 7.5, 150 mM NaCl, 2 mM EDTA) and stored at  -80 °C. Cross-linked nuclei were then subjected to ChIP analysis. For ChIP-seq, ChIP DNA was ligated to single-end sequencing adapters and amplified. DNA fragments of 300-800 bp in size were purified and sequenced (50 base) on an Illumina Hi-seq2000 instrument (Illumina, California, USA).
attributes.tag: GEO Accession
attributes.value: GSM1301669