Double Digest Guide
From 2006.igem.org
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| - | + | ===iGEM Double Digest Guide=== by Karmella Haynes, 2006 | |
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| - | + | ===References=== | |
| - | * [https://dspace.mit.edu/handle/1721.1/21168 | + | * [https://dspace.mit.edu/handle/1721.1/21168:Knight, Tom. Idempotent Vector Design for Standard Assembly of Biobricks] |
Revision as of 23:44, 19 September 2006
===iGEM Double Digest Guide=== by Karmella Haynes, 2006
| Enzymes | Buffer | Temperature | Purpose |
| EcoRI, XbaI | Low | 37°C | To create a "Front Vector" |
| EcoRI, SpeI | Low | 37°C | To create a "Front Insert" |
| SpeI, PstI | Medium | 37°C | To create a "Back Vector" |
| XbaI, PstI | Low | 37°C | To create a "Back Insert" |
| EcoRI, PstI | Promega® Buffer H | 37°C | To excise entire insert or validate part size |
Davidson Buffers [10 mM Tris-HCl pH 7.5, 10 mM MgCl2, 0.1 mg/mL BSA, X mM NaCl]
- 0 (zero), 0 NaCl
- Low, 50 mM NaCl
- Medium, 100 mM NaCl
- High, 150 mM NaCl
Promega® Buffer H [90 mM Tris-HCl pH 7.5, 10 mM MgCl2, 50 mM NaCl]
