I performed a digestion that ended up at midnight, but forgot to inactivate the enzymes (HaeIII and RSAI) before freezing the products. I plan to run a gel to differentiate yeast isolates. Can I just run the gel or should I inactivate the enzymes after defrosting the samples?
Kindly check https://www.addgene.org/protocols/restriction-digest/
Also check https://www.takarabio.com/learning-centers/cloning/traditional-molecular-cloning/restriction-enzyme-overview/inactivation-of-restriction-enzymes
There appear to be two reasons for enzyme inactivation prior to freezing the product. Firstly, there is water in the product, so during freezing there may be a temporary increase in the rate of reactions catalyzed by the enzymes used. Secondly, freezing will significantly reduce the rate of enzymatic reactions, but after defrosting, their rate will increase again.
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