Matt Gethers/CRI, Thailand/Labwork/Digests/NdeI and ApaI Double Digest
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NdeI and SacI Double Digest
Reaction Mix
| Reagent | Volume |
| Template | 10 μl |
| NdeI | 0.5 μl |
| SacI | 0.5 μl |
| Fast Digest Buffer | 2 μl |
| H20 | 7.0 μl |
| Total | 20 μl |
Reaction Conditions
Incubate mixture at 37oC for 30 minutes and inactivate at 65oC for 15 minutes on the heat block.
Run Notes
7.29.08
Used to digest pCM351 to excise Lox/Gentamcin cassette. 2.5 μl pCM351, 0.5 μl both enzymes, 2 μl buffer, and 14.5 μl water. Incubated at 37 degrees for 1 hour, 15 minutes, inactivated at 65 degrees for 15 minutes. Ran out on gel and excised ~1 Kb band. Did a gel purification on the morning of 7.30.08 and eluted in 50 μl elution buffer.
8.12.08
Digested pCM351 to excise Lox/Gentamcin cassette. 2.5 μl pCM351, 0.5 μl both enzymes, 2 μl buffer, and 14.5 μl water. Incubated at 37 degrees for 30 minutes, inactivated at 65 degrees for 15 minutes. Ran out on gel and excised ~1 Kb band. Did a gel purification and eluted in 50 μl elution buffer.
