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Team:USTC-China/results
From 2012.igem.org
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Revision as of 14:19, 23 September 2012
RESULTS
DNA digestion
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)
Double Digestion
| Volume (μl) | |
| Plasmid DNA | Up to 1μg |
| 10×FastDigest Buffer | 2 |
| Enzyme1 | 1 |
| Enzyme2 | 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)
