Restriction Digestion Protocol
Check the compatibility of two restriction enzymes and decide which digestion buffer to use (look at the NEB catalog)
Double digest (digest with 2 enzymes at once)
| Vol (uL) | Reagent |
| 16 | ddH2O |
| 7.5 | Digestion Buffer 10X |
| 7.5 | BSA 10X |
| 40 | DNA |
| 2 | Enzyme 1 |
| 2 | Enzyme 2 |
| 75 ul Total | |
| Incubate at 37C for 2 hours | |
For single digest: alter recipe, perform digest with one enzyme
Then perform PCR cleanup procedure, elute with 40 uL
Then can digest DNA with second enzyme