Right now, the digest protocol tries to make the reaction volume as small as possible. Specifically, no water is added unless it is needed to get the amount of enzyme below 10% of the reaction. I can't exactly say why, but I'm not really happy with this approach. Even though it minimizes volume, it just doesn't jive with how most reactions work.
However, NEB also recommends using 50 µL per 1 µg of DNA. Note that isn't a precise definition, because the number of sites per microgram isn't specified. Also, it's possible for the input DNA to be dilute enough that 1 µg would be more than 50 µL. Still, it suggests an alternative way to setup these reactions. I could still have an option for minimizing volume.
Right now, the digest protocol tries to make the reaction volume as small as possible. Specifically, no water is added unless it is needed to get the amount of enzyme below 10% of the reaction. I can't exactly say why, but I'm not really happy with this approach. Even though it minimizes volume, it just doesn't jive with how most reactions work.
However, NEB also recommends using 50 µL per 1 µg of DNA. Note that isn't a precise definition, because the number of sites per microgram isn't specified. Also, it's possible for the input DNA to be dilute enough that 1 µg would be more than 50 µL. Still, it suggests an alternative way to setup these reactions. I could still have an option for minimizing volume.