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Team:Groningen/protocols/Digestion
From 2013.igem.org
Digestion
Materials:
- H2O
- 1.5ml tubes
- Ligation buffer
- Restriction enzymes
- DNA
Reaction preparation:
| Component | 50μl reaction | Final concentration |
| H2O | up to 50μl | |
| 5x Phusion Buffer | 10 | 1x |
| 10mM dNTPs | 1 | 200μM |
| F-primer | 1 | 0.2μM |
| R-primer | 1 | 0.2μM |
| Template DNA | 1μl | ~1ng |
| Phusion Pol. | 0.5μl | 0.02U/μl |
Cycling instructions:
| Temperature | Time | Number of cycles |
| 98°C | 30sec | 1 |
| 98°C | 10sec | 30 |
| primer annealing temperature | 30sec | 30 |
| 72°C | 30sec/kb | 30 |
| 72°C | 10min | 1 |
| 4°C | ∞ | 1 |
iGEM 2013 Groningen
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