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BsaWI > FAQ |  | BsaWI FAQ
See the Restriction Endonucleases FAQ also.
Q1: The NEB catalog has historically stated that activity in NEBuffer 4 is less than 100% yet this enzyme is now supplied with NEBuffer 4. What have you changed?
Q2: Has the conversion to NEBuffer 4 altered any of the properties of the restriction enzyme?
Q3: If I have an old tube of enzyme, what NEBuffer should I use?
Q4: Will the new enzyme work in the originally supplied NEBuffer?
Q5: Why is NEB switching this restriction enzyme to NEBuffer 4?
Q6: Do degenerate recognition sites need to be palindromic?
Q1: The NEB catalog has historically stated that activity in NEBuffer 4 is less than 100% yet this enzyme is now supplied with NEBuffer 4. What have you changed?
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: Do degenerate recognition sites need to be palindromic?
A6: Most restriction enzyme recognition sites are palindromic and include only specified base pairs (i.e., EcoRI recognizes GAATTC). However, some enzymes have degenerate sites, meaning that they contain one or more base pairs that are not specifically defined (i.e., BsrFI recognizes RCCGGY, where R= A or G and Y= C or T). For degenerate enzymes, any base represented by the single letter code may be present at either location in the recognition site for cleavage to occur. For example, BsrFI recognizes all of the following sequences: ACCGGC, ACCGGT, GCCGGC, GCCGGT.
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