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We have observed that dose response curves vary for different guide sequences, depending on potency of the guide RNA. We recommend titrating the amount of ribonucleoprotein (RNP) and keeping the amount of electroporation enhancer fixed.
For the Nucleofector™ System (Lonza), we recommend 3 µM of Alt-R Cas12a Electroporation Enhancer, and for the Neon™ System (Thermo Fisher Scientific), we recommend 1.8 µM of Alt-R Cas12a Electroporation Enhancer. Toxicity may be observed at high concentrations of enhancer.