GSM2406793: WT_NSCs_H3K4me1_ChIPSeq; Mus musculus; ChIP-Seq
For each line, neurospheres were dissociated and 4 million single cells were crosslinked with 1% formaldehyde for 10 minutes at room temperature. Reaction was quenched with 0.125 M Glycine, cells were washed with cold PBS and lysed according to Vermunt et al., 2016. Nuclei pellets were resuspended in 160μl sonication buffer and divided over two microtubes for shearing in the Covaris S series with the following settings for 12 cycles of 60 seconds: intensity 3, duty cycle 20%, 200 cycles/bursts.  Chromatin immunoprecipitation steps after sonication were performed as described previously (Vermunt et al., 2016) using 50 μl DynaI protein G beads that were preincubated with 5 μg Ab4729 (Abcam) for H3K27ac or 5 μg Ab8894 (Abcam) for H3K4me1. Whole cell extract of 4 million cells was split onto both antibodies, resulting in the use of 2 million cells per ChIP. Libraries were made using the Illumina Truseq DNA library protocol and sequencing was done at the MIT BioMicro Center (http://openwetware.org/wiki/BioMicroCenter).
GEO Accession
GSM2406793