New England Biolabs provides a color-coded 10X NEBuffer with every restriction endonuclease to ensure 100% activity. Some enzymes require an NEBuffer which is different from the four standard NEBuffers (1, 2, 3 or 4) for optimal activity. To help select the best conditions for double digests with these enzymes that require a unique (designated “U”) NEBuffer, the table below lists the activity of the most commonly used restriction endonucleases in the unique NEBuffers supplied with the following enzymes: BsrFI, DpnII, EcoRI and SspI.
Performing double digests with enzymes supplied with a unique NEBuffer and not one of the four standard NEBuffers is usually simple. In most cases, double digests using any of these enzymes with an enzyme that is supplied with a standard NEBuffers can be done in the unique NEBuffer. This will ensure that the enzyme with the more specific buffer requirements will work optimally. When using restriction endonucleases in non-optimal NEBuffers, more enzyme or longer digestion time may be needed to compensate for the slower rate of cleavage under those conditions.
If BSA or SAM is supplied with an enzyme, it is included in all NEBuffer activity reactions.
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NEBuffer Activity | Double Digestion | Time-Saver Enzymes | High Fidelity (HF) Restriction Enzymes
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