FOR FILLING IN 5' OVERHANGS
DNA Polymerase I (E. coli), Large Klenow Fragment, Klenow Fragment (3'-5' exo-), and T4 DNA polymerase are all active in all NEBuffers, but NEBuffer 2/2.1/r2.1 provides optimal activity. Optimal activity of T7 DNA Polymerase is seen in its own buffer, and it has 50% reduced activity in the other NEBuffers.
For comparisons of NEBuffer compositions, click here. To see the activity of more DNA modifying enzymes in rCutSmart Buffer, click here.
FOR BLUNTING OVERHANGS
| NEBuffers | DNA Polymerase I (E. coli) | DNA Polymerase I, Large (Klenow) Fragment | T4 DNA Polymerase | T7 DNA Polymerase |
|---|---|---|---|---|
| 1 | Y | N | Y | 50% |
| 1.1 | Y | N | Y | 50% |
| r1.1 | Y | N | Y | 50% |
| 2 | Optimal | Optimal | Y | 50% |
| 2.1 | Y | Y | Y | 50% |
| r2.1 | Y | Y | Optimal | 50% |
| 3 | Y | Y | N | 50% |
| 3.1 | Y | Y | N | 50% |
| r3.1 | Y | Y | N | 50% |
| 4 | Y | Y | Y | 50% |
| CutSmart | Y | Y | Y | 50% |
| rCutSmart | Y | Y | Y | 50% |
| T4 DNA Ligase Buffer | Y | Y | ||
| T7 DNA Polymerse Reaction Buffer | Optimal |
"Y" means yes, this polymerase can be used in the specified buffer. "N" means no, this polymerase cannot be used in the specified buffer. Optimal buffers are indicated. Klenow (3'-5' exo-) cannot be used to remove overhangs.