Unique NEBuffer Chart
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: BamH I, BceA I, BsrF I, Dpn II, EcoR I, Sau3A I and Ssp I.
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. Additional double digestion information can be found on the previous page.
Note: The values listed in this table are approximate. They were obtained using each enzyme's specific unit assay substrate DNA.NEBuffer Chart | All NEB Buffers
Click for the Chart Legend.
| Enzyme | Supplied NEBuffer | % Activity in Unique NEBuffers | |||
| BamH
I BceA I |
Dpn II | EcoR
I Ssp I BsrF I |
Sau3A I | ||
| Aat II | 4 | 0 | 0 | 25 | 0 |
| Avr II | 2 | 100 | 100 | 100 | 100 |
| BamH I | Udd + BSA | 100 | 100 | 100 | 100 |
| Bgl II | 3 | 50 | 0 | 50 | 25 |
| Bsg I | 4 + SAM | 10 | 10 | 10 | 25 |
| Eag I | 3 | 100 | 10 | 100 | 25 |
| EcoR I | Udd | 50 | 0 | 100 | 10 |
| EcoR V | 3 + BSA | 100 | 0 | 100 | 50 |
| Hind III | 2 | 10 | 0 | 50 | 0 |
| Kpn I | 1 + BSA | 10 | 10 | 25 | 25 |
| Mse I | 2 + BSA | 50 | 10 | 50 | 50 |
| Nco I | 4 | 25 | 10 | 100 | 75 |
| Nde I | 4 | 100 | 0 | 100 | 50 |
| Nhe I | 2 + BSA | 10 | 50 | 75 | 25 |
| Not I | 3 + BSA | 25 | 0 | 100 | 25 |
| Pst I | 3 + BSA | 50 | 0 | 50 | 10 |
| Pvu I | 3 + BSA | 50 | 0 | 75 | 10 |
| Sac I | 1 + BSA | 10 | 0 | 25 | 10 |
| Sac II | 4 | 100 | 0 | 100 | 25 |
| Sal I | 3 + BSA | 100 | 0 | 100 | 25 |
| Sma I | 4 | 0 | 0 | 0 | 0 |
| Spe I | 2 + BSA | 50 | 25 | 75 | 50 |
| Sph I | 2 | 10 | 10 | 50 | 25 |
| Xba I | 2 + BSA | 0 | 10 | 10 | 100 |
| Xho I | 2 + BSA | 100 | 0 | 100 | 50 |
| Xmn I | 2 + BSA | 10 | 0 | 10 | 10 |