Restriction
Enzymes
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.