I am using CRISPR Knockout and activation products from Santacruz biotechnology, particularly double Nickase Plasmid. I followed their protocol and tried a different ratio of DNA plasmid and transfection reagent, 1/10, 2/10, 1/5, 2/5 for 24, and 48 hours. Unfortunately, the cells die in all the trials. In the beginning, the cells look like they contain granules and start detachment.