A1: All our restriction enzymes were re-assayed in both NEBuffer 4 vs. the previously supplied NEBuffer. In a few cases minor formulation changes were required to bring the activity to 100% in NEBuffer 4.
Q2: Has the conversion to NEBuffer 4 altered any of the properties of the restriction enzyme?
A2: In general the properties of the restriction enzyme remain the same although for some enzymes, some minor changes in heat inactivation, Time-Saver™ qualification, etc. were observed. All the updated information can be found on the supplied data card as well as at www.neb.com.
Q3: If I have an old tube of enzyme, what NEBuffer should I use?
A3: Our restriction enzymes are color coded on the label for the appropriate NEBuffer and can either be used with the previously supplied NEBuffer or with NEBuffer 4. To ensure 100% cleavage in NEBuffer 4, additional units of enzyme and/or longer incubation time may be necessary.
Q4: Will the new enzyme work in the originally supplied NEBuffer?
A4: Yes it will. In all instances the restriction enzyme will have 100% activity in the originally supplied NEBuffer.
Q5: Why is NEB switching this restriction enzyme to NEBuffer 4?
A5: Our main goal is to simplify the NEBuffer system so that the majority of restriction enzymes are compatible in a single buffer. We now supply 162 restriction enzymes with NEBuffer 4 including all the new High Fidelity (HF) restriction enzymes.
Q6: Is MboII affected by methylation?
A6: MboII is blocked by overlapping dam methylation.
Q7: Why is MboII cut DNA difficult to ligate?
A7: MboII produces DNA fragments that have a single-base 3´ extension which are more difficult to ligate than blunt ended fragments.
Q8: Is extended digestion of MboII recommended?
A8: Extended digestion is not beneficial because the enzyme is not stable in reaction.
