FAQ: How do I dilute the enzyme to 1 μM for in vitro reactions?

If planning to use higher concentration EnGen Lba Cas12a (NEB #M0653T) for in vitro digestion of DNA, the enzyme can be diluted to 1 μM in 1X Buffer 2.1 (NEB #B7202) and used immediately. If the 1 μM dilution will be stored at -20°C, it should be diluted in the enzyme storage buffer: 500 mM NaCl, 20 mM sodium acetate, 0.1 mM EDTA, 0.1 mM TCEP and 50% glycerol (pH 6.0 @ 25°C).

When diluting 100 µM EnGen Lba Cas12a (Cpf1) for use in in vitro cleavage assays, supplementing reactions with 10 mM DTT can in some cases improve substrate cleavage.