GSM9336977: MEF cells treated with Alexidine (Z22 ChIP); Mus musculus; ChIP-Seq
Briefly, the cells were fixed for 10 min with 1% (w/v) formaldehyde in DMEM. Following this, the crosslinked DNA was sheared with an enzymatic shearing cocktail (provided in the kit) for 10 min to aid digestion of chromatin, yielding DNA of about 200-500 bp in length. Further, 10 mg of the sheared DNA was immunoprecipitated with 2 mg of anti-Z-DNA (Z22) antibody overnight at 4°C. The construction of library was performed using NEBNext® UltraTM DNA Library Prep Kit for Illumina (New England Biolabs, UK) according to the manufacturer's instructions. Briefly, the chipped DNA was ligated with adaptors. After purification, PCR reaction was done with adaptor-ligated DNA and index primer for multiplexing sequencing. Library was purified by using magnetic beads to remove all reaction components. The size of library was assessed by Agilent 2100 bioanalyzer (Agilent Technologies, Amstelveen, The Netherlands). High-throughput sequencing was performed as paired-end 100bp sequencing using NovaSeq 6000 (Illumina, Inc., USA).