ChIP-Atlas: SRX1733609

Antigen

Cell type Class: Blood
Cell type: Bone Marrow Cells
MeSH Description: Cells contained in the bone marrow including fat cells (see ADIPOCYTES); STROMAL CELLS; MEGAKARYOCYTES; and the immediate precursors of most blood cells.

library_strategy: ChIP-Seq
library_source: GENOMIC
library_selection: ChIP
library_construction_protocol: 2 million cells were crosslinked with 1% formaldehyde at room temperature for 10 minutes. Crosslinking was terminated by addition of 0.125M glycine and washed with ice-cold PBS containing protease inhibitor cocktail (PIC, Roche). Samples were sonicated to 200-500bp fragments for 8 cycles at 5 minutes each (Biorupter, Diagenode). Sheared chromatin were incubated with FLI1 antibody (ab15289;Abcam) and Protein A (Dynabeads, Invitrogen). Chromatin were reverse crosslinked at 68°C for 3 hours at mixed in Elution buffer (1M Tris pH 7.5, 0.5M EDTA, 5M NaCl, 10% SDS, Proteinase K) at 1100rpm. DNA was purified by MiniElute PCR Purification Kit (Qiagen) and eluted in 22μL EB buffer. Purified DNA was used to generate libraries with the ThruPLEX-FD preparation kit (Rubicon)