Fetal heart 4: H3K27me3 human cardiac myocyte nuclei
Isolated cardiac myocyte nuclei were fixed with paraformaldehyde, lysed and sonicated using a Bioruptor (Diagenode) with 30 cycles (30 s on/ 30 s off; high energy). The isolated chromatin was used for the ChIP experiment. The immunoprecipitation was performed with 4  g of anti-H3K27me3 antibody (Millipore, 07-449) overnight. Reversal of cross-linking and DNA purification were performed using the ChIP-IT High Sensitivity Kit (Active Motif) following the manufacturer's manual. The eluted DNA was concentrated by evaporation at room temperature using a vacuum concentrator (Eppendorf) to 55.5  l. Libraries were prepared using the NEBNext Ultra DNA Library Prep Kit for Illumina (NEB) according to the manufacturer's manual omitting the size selection.
SUB3380142
Fe4_H3K27me3