Chromatin opening on CBFA2T3::GLIS2 iPSC-derived progenitors
Chromatin opening on CBFA2T3::GLIS2 iPSC-derived progenitors
CD41+ Flow sorted differentiation day-13 hematopoietic cells from ctrl or EG iPSC, leukemic NSG mice or patient samples were collected. 5x104-1x105 GFP+ live-sorted cells were spun at 500g for 5 min, washed with cold DPBS, lysed in cold lysis buffer (10 mM Tris-HCl (pH 7.4), 10 mM NaCl, 3 mM MgCl2, and 0.1% NP40, 3min on ice) and immediately spun at 500g for 10 min. The pelleted nuclei were resuspended in 50 µL of transposase reaction mix (Tagment DNA TDE1 Enzyme and Buffer Kit, Illumina) for 30 min at 39°C.  PCR purification was performed using Agencourt AMPure XP magnetic beads (Beckman Coulter A63880) in order to remove large fragments and remaining primers. Library quality was assessed using an Agilent 2100 Bioanalyzer using a High Sensitivity DNA chip (Agilent Technologies 5067-4626). Librairies were constructed by PCR using the Nextera DNA CD index kit (Illumina) and the NEBNext Ultra II Q5 master mix (NEB).