title: GSM8006448: CMSIP-Ogt_iKO mESC, +4OHT 6-days, rep#2; Mus musculus; OTHER
library_name: GSM8006448
library_construction_protocol: In order to ensure a high purity of our samples, we isolate specifically the YFP+ population (in the case of 4-OHT-treated Ogt iKO mESC) at the same time that eliminating the presence of mitomycin-C-treated MEFs (used as feeders during our mESC cultures) by FACS (Fluorescence-Activated Cell Sorting) using a FACSAria cell sorter (BD Biosciences). The dissociated mESC were first labeled with the eBioscienceTM Fixable Viability Dye eFluorTM 780 (ThermoFisher, 65- 0865-14) to select viable cells and then stained for surface markers with indicated fluorochrome-conjugated antibodies diluted in FACS buffer (PBS 1X, 0.5% BSA, 0.01% NaN3): CD90.2 (Biolegend, 140319) to positively distinguish the mESC and CD326 (Biolegend, 118227) to negatively exclude MEFs. CMS-IP with spike ins