Smolke:Reaction times

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Digests NEB says 5 minutes for most of its enzymes. If you're gel-purifying out an insert, 5-10 minutes seems to be sufficient. If you want to be more certain (if you need the vector, for example, and the insert is too small to distinguish singly from doubly cut), a hour or two seems fine (Jkm) Ligations NEB says 5 minutes for its quick ligase and 10 minutes for its standard ligase. 30 minutes is more than sufficient (Jkm)