Runx1-ires-GFP+/Gata1::mCherry+ cells from embryoid bodies after 5 days of differentiation of ES cells towards blood
chip antibody
rabbit anti-Gata1, Abcam ab11963, lot I0303
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Approximately 7x106 FACS-sorted day 5 EB cells per ChIP were cross-linked using formaldehyde to a final concentration of 1%. As samples were pooled from several sorts, isolated nuclei were frozen on dry ice cold isopropanol and stored at -80°C. During the immunoprecipitation step, 4 μl recombinant histone 2B (New England Biolabs) and 1 μl of mouse RNA (Qiagen; diluted 1/5 in IP dilution buffer) were added as carriers, followed by 7 μg of primary antibody Sequencing libraries were prepared using the TruSeq Kit (Illumina) for high throughput sequencing on an Illumina HiSeq 2500, according to manufacturer’s instructions, with size selection for fragments of 150-400 bp.