A1: BSA is always supplied with enzymes that require it, as well as with each of our NEBuffer 1, 2, 3 and 4 packs and unique NEBuffer packs that are for enzymes requiring BSA. It can also be ordered separately as NEB# B9001S (25.0 mg).
Q2: Does NEB supply reaction buffers with their restriction enzymes?
A2: New England Biolabs provides a color-coded 10X NEBuffer with each restriction enzyme to ensure optimal (100%) activity. The buffer should be used at 1X concentration in the reaction. Some restriction enzymes require bovine serum albumin (BSA) at a final concentration of 100 μg/ml for optimal activity. When required, BSA is supplied as a 10 mg/ml (100X) stock and should be added to the reaction mixture.
Q3: When setting up a restriction digest, if BSA is not listed as a reaction component, can it have a negative effect if added?
A3: Restriction enzymes that do not require BSA for optimal activity are not adversely affected if BSA is present in the reaction.
Q4: Why do I need to add BSA to my restriction digest?
A4: Adding BSA to a reaction lessens enzyme loss on tube and pipette tip surfaces. BSA stabilizes enzymes in reaction. The stabilizing effects are most pronounced in overnight reactions (Robinson D. unpublished observation).
Q5: The buffer arrived thawed; is it and the enzyme still active?
A5: The buffers will often thaw when shipped on wet ice. This is often due to the high salt concentration of the 10X buffer. If the ice pack is still solid and the contents are cold to the touch the buffer and enzyme are fine.
Q6: Is SAM supplied with enzymes that require it in the reaction buffer?
A6: SAM is provided with all enzymes that require it. It can also be ordered separately as product NEB# B9003S (5 vials of 100 μl each of a 32 mM solution).
