Single Digestion
1.Performed with Thermo Scientific FastDigest. There are four restriction endonucleases we used in our experiments: EcoRI, XbaI, SpeI, PstI.
Single Digestion
|
Volume (μl) |
| Plasmid DNA |
Up to 1μg |
| 10×FastDigest Buffer |
2 |
| Enzyme |
1 |
| ddH2O |
To 20 |
| Total |
20 |
2. Mix gently and spin down.
3. Incubate at 37°C in a heat block or water thermostat for 30 min.
4. Inactivate the enzyme by adding glycerol DNA loading buffer, or by heating for 5 min at 80°C(only for EcoRI) or 20min at 65°C(only for XbaI)
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