GSM4551211: input_uninfected_H3K27ac_1; Mus musculus; ChIP-Seq
Chromatin was immunoprecipitated using the True MicroChIP kit (Diagenode) with a modified protocol: cells were lysed before chromatin shearing using a Bioruptor Pico Sonicator (Diagenode; 5 cycles: 30 seconds ON - 30 seconds OFF; fragments 100 - 300 bp). 10 % of sheared chromatin was kept as input control and 90 % was immunoprecipitated using antibodies against either H3K27ac, H3K4me1 or H3K9me (all Diagenode) over night at 4 ̊C. The next morning Protein A coated beads (Diagenode) were added and, following a 6 hour incubation at 4 ̊C, unbound chromatin fragments were removed by thorough washing. ChIP and input DNA was then decrosslinked from proteins and purified using MicroChip DiaPure columns (Diagenode). Sequencing libraries were prepared using the MicroPlex v2 Library preparation kit (Diagenode) according to manufacturers instructions. Library amplification was monitored in real time using a Lightcycler 480 (Roche) to determine optimal cycle number. Amplified libraries were quantified using Qubit (dsDNA HS assay) and fragment quality and size distribution assessed by Bioanalyser (DNA HS Kit). Following equimolar pooling, libraries were purified using AMPure XP beads and quantity and quality of the library pool confirmed. Pooled libraries were sequenced by Edinburg Genomics, University of Edinburgh (UK) on HiSeq 4000 (75 bp paired end reads) or NovaSeq S1 (100 bp paired end reads, both Illumina).
GEO Accession
GSM4551211