GSM2113514: SHEP21_2HR_INPUT_CHIPRX; Homo sapiens; ChIP-Seq
ChIP-Rx was performed as described previously substituting mouse embryonic stem cells in place of Drosophila S2 cells (Orlando et al., 2014). Mouse embryonic stem (ES) cells were grown to 75% confluence and cross-linked with 1% formaldehyde followed by quenching (125 mM glycine) as described in the previous section. Cells were washed in cold PBS and harvested by cell scraper in cold PBS with protease inhibitors (Roche). Cells were centrifuged at 1650 x g for 5 minutes and flash frozen and stored at -80°C 10E06 cells per pellet. Fixed mouse ES cell pellets were resuspended in a cytosolic lysis buffer and then spiked directly into the cytosolic lysate of fixed neuroblastoma cells at a ratio of 5% of the total number of cells. The lysate (neuroblastoma cells spiked with mouse ES cells) was then carried through the ChIP protocol. Libraries were prepared using the Rubicon Thruplex FD library preparation kit (cat#: 400427) per manufacturers instructions.
GEO Accession
GSM2113514