Component | Amount | Final |
---|---|---|
5× ligase reaction buffer | 4 µl | 1× |
ScaI → XmaI DNA fragment ends | 30–120 fmol | 30–120 fmol |
BspEI → ScaI DNA fragment ends | 30–120 fmol | 30–120 fmol |
T4 DNA ligase (1 U µl−1) | 1 µl | 1 U |
Nuclease-free water | Up to 20 µl |
Component | Amount | Final |
---|---|---|
5× ligase reaction buffer | 4 µl | 1× |
ScaI → XmaI DNA fragment ends | 30–120 fmol | 30–120 fmol |
BspEI → ScaI DNA fragment ends | 30–120 fmol | 30–120 fmol |
T4 DNA ligase (1 U µl−1) | 1 µl | 1 U |
Nuclease-free water | Up to 20 µl |