GSM3478664: H3K4me1 ChIPSeq Tet2-/-,Rep2; Mus musculus; ChIP-Seq
The nuclei/chromatin suspension was sonicated using a Bioruptor Pico Sonnicator (Diagenode) for 20 cycles at 4°C. Sheared chromatin size was confirmed to be between 100-800 base-pairs on average by agarose gel. Two million cell equivalents were used for each ChIP. The following antibodies were used: H3K4me1 (Abcam, ab8895), H3K27ac (Abcam, ab4729), PU.1 (Santa Cruz Biotechnology, sc-352), Erg-1/2/3 (Santa Cruz Biotechnology, sc-354) and antibodies against total Stat5 (Santa Cruz, sc-835). ChIP samples were quantified using the Qubit Fluorometric quantification System (ThermoFisher) and using a DNA high sensitivity DNA kit on a Bioanalyzer (Agilent). ChIP-seq libraries were made using the Multiplex Library Preparation Kit (Diagenode). Nine cycles of PCR were used for all libraries. After library DNA was purified, library concentration was measured by Qubit, and size distribution was determined by Bioanalyzer. Each ChIP-seq library was sequenced for single-end reads of Illumina HiSeq 4000 (between 100-800 bp read length). Biological replicates for ChIP-seq were performed on independently derived primary mast cell cultures from 2 different mice for each sample and both sequenced.
GEO Accession
GSM3478664