title: GSM6568235: ESC input replicate1 [ChIP-Seq]; Mus musculus; ChIP-Seq
library_name: GSM6568235
library_construction_protocol: Cell pellets were washed twice in PBS and resuspended in 300ul lysis buffer (10mM Tris pH8, 1mM EDTA, 0.5% SDS) for at least 15 minutes.  Next, chromatin was sonicated in a Pico bioruptor device for 10 cycles with the length of the intervals being 30sec on/off, in order to produce 300-800 bp chromatin fragments. Sonicated chromatin was then spun down for 15 minutes at 4°C at 22.000g and 10μl of the sheared soluble chromatin solution was used in order to check the shearing efficiency and the rest was kept at 4oC. 5% of each sample was kept as an input while the rest of the supernatants were diluted 5 times with dilution buffer (0.01% SDS, 1.1% triton,1.2mM EDTA,16.7mM Tris pH8, 167mM NaCl) and incubated with 3μg H3K27ac antibody (ab4729) O/N under agitation at 4°C. 25ng of immunoprecipitated material and input were used for ChIP-seq library preparation using the KAPA Hyper prep kit (KK8502) according to manufacturer's instructions.