Hi all, I used the Thermo-ANZA series for overnight digestion of plasmid backbone to ensure clean digestion without any problem.
How about NEB? The website said all NEB "time-saver enzyme" series can have fast and withstand overnight digestion. Did anyone encounter any problem with overnight digestion with these enzymes?
No I haven't had any problems with ON digestions. I once did not have the time to purify the plasmid digest the next morning, so I kept the digest at 4°C until the evening. Also no problems. NEB kinda rocks
In general NEB prolonged digestion is not a problem. Especially the HF one is suitable for longer digestions since they have no star activity (which is one of the disadvantages of prolonged incubation). This is a good site to check if prolonged incubation makes sense because some enzymes are not active after some time
https://international.neb.com/tools-and-resources/usage-guidelines/restriction-endonucleases-survival-in-a-reaction
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