Restriction enzymes recognize specific, short sequences of double-stranded DNA where they then cleave it. For more information, see any product usage sheet that accompanies a particular enzyme.
Following is a general protocol for setting up a restriction digest:
| 10X buffer | 1.5 ul |
| BSA (10ug/ul) | 0.15ul |
| DNA | 3.0ul |
| ddH2O | to 14.5ul |
*If digesting more than one sample,
Mix, then add:
| restriction enzyme | 0.5ul |
| Final Volume | 15.0ul |
Mix gently. Incubate in 37°C water bath (or appropriate temperature) for 1-4hours.
Check on an agarose gel.