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Team:UC Davis/Protocols
From 2011.igem.org
Contents |
Restriction Enzyme Double Digest
Materials
- - 22 uL dH2O
- - 1 uL BSA
- - 5 uL Buffer
- - 20 uL Template
- - 1 uL Enzyme 1
- - 1 uL Enzyme 2
Buffer Compatibility Chart
| 1 | 2 | 3 | 4 | |
|---|---|---|---|---|
| EcoRI | 100 | 100 | 100 | 100 |
| SpeI | 75 | 100 | 25 | 100 |
| PstI | 75 | 75 | 100 | 50 |
| NheI | 100 | 100 | 10 | 100 |
| XbaI | 0 | 100 | 75 | 100 |
Procedure
- - Mix reactants thoroughly.
- - Place at 37 C for 3 hours.
- - Increase to 80 C for 20 minutes to kill enzymes (some enzymes need only a 65 C heatkill, check enzyme).
- - Run on a gel and extract product.
