GSM4403876: DM.HSPC_H3K4me3_rep2; Mus musculus; ChIP-Seq
Chromatin immunoprecipitation was performed with iDeal ChIP-seq kit for Histones (Diagenode). Briefly, 1 x 106 Lin- HSPCs or neutrophils were crosslinked with formaldehyde at a final concentration of 1% for 10 minutes and then quenched by addition of Glycine for 5 minutes incubation. Lysis buffers iL1 and iL2 were used to prepare the nuclei pellets. Chromatin was sheared in Shearing buffer iS1 using the Bioruptor Plus (Diagenode) for 20 cycles (each cycle for 30 seconds “ON” and 45 seconds “OFF”) at high power setting. Immunoprecipitation, wash, decrosslinking and elution were carried out as per the manufacturer's protocol, using 1 µg antibody. 10% of the sheared chromatins were kept aside as input samples. Immunoprecipitated DNA or input DNA were used library preparation by using TruSeq ChIP Sample Prep Kit (Illumina), following standard procedures from manufacturer. The library DNA was quantified using KAPA Library Quantification Kit (Kapa Biosystems) and library average size was determined by Agilent DNA 1000 Kit (Agilent Technologies).
GEO
GSM4403876