Double Digest Guide
From 2006.igem.org
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'''iGEM Double Digest Guide''' by Karmella Haynes, 2006 | '''iGEM Double Digest Guide''' by Karmella Haynes, 2006 | ||
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'''Promega® Buffer H''' [90 mM Tris-HCl pH 7.5, 10 mM MgCl2, 50 mM NaCl] | '''Promega® Buffer H''' [90 mM Tris-HCl pH 7.5, 10 mM MgCl2, 50 mM NaCl] | ||
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| + | '''References''' | ||
| + | * [https://dspace.mit.edu/handle/1721.1/21168| Knight, Tom. Idempotent Vector Design for Standard Assembly of Biobricks] | ||
Revision as of 23:42, 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]
References
