▲ CRITICAL STEP Add D10 very slowly to the side of the well, and do not use cold medium, as cells can detach easily. Puromycin selection can be applied at a concentration of 1–3 μg ml–1 for HEK 293FT cells (may vary depending on the cell line). 13Incubate the cells for a total of 48–72 h after transfection before passaging them for downstream applications or harvesting for indel analysis.
