Instrument: Illumina HiSeq 2000
Strategy: ChIP-Seq
Source: GENOMIC
Selection: ChIP
Layout: SINGLE
Construction protocol: Adult mouse liver were collected at 6 different time of the day (mouse held under LD12:12, time points every 4hrs starting at ZT2) Mouse liver were crosslinked with 1% formaldehyde for 10min, and nuclei were purified using standard sucrose cushion protocol. Nuclei were resuspended, flash frozen in liquid nitrogen and stored at -80C. Nuclei were thawed on ice, pelleted down and chromatin either was digested with micrococcal nuclease or sonicated using Bioruptor. Mnase digestion was performed for 15min using 30,000 gel unit per reaction Following chromatin shearing, chromatin was imunoprecipitated using an anti-CLK antibody. Immunoprecipitated chromatin and inpur fraction was purified and used for generating illumina libraries Illumina libraries were generated using standard protocol: end repair, add A, adapter ligation, amplification, gel purification.