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We see dose response curves that vary for different guide sequences, depending on the potency of the guide sequence. We recommend titrating the amount of RNP for each cell type of interest.
Start with a 1:1 ratio of enhancer:RNP and optimize using a titration gradient of the RNP. For the Amaxa® Nucleofector® System (Lonza), we recommend 4 µM of Alt-R® Cas9 Electroporation Enhancer, and for the Neon® System (Thermo Fisher Scientific), we recommend 1.8 µM of Alt-R Cas9 Electroporation Enhancer.