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Template:Team:KULeuven/26 August 2009/VanillinProduction
From 2009.igem.org
- Miniprep of fluid cultures
Nanodrop results:
| Part | concentration (ng/μl) | 260/280 λ |
|---|---|---|
| SamS I | 53,9 | 1,87 |
| SamS II | 39,4 | 2,16 |
| SamS III | 57,1 | 2,07 |
| EF I | 76,8 | 1,94 |
| EF II | 83,4 | 2,04 |
| EF III | 61,5 | 2,01 |
- Restriction:
-We performed 2 simultaneous digests: one to verify the length of the product (Restriction2Test) using SAMS II and EF III and another to prepare for ligation (Restriction4Real) with promotor and terminator using SAMS III and EF II
-We cut with the 2 enzymes separately, allowing a one hour incubation in between. EF: E/S SAMS: E/S
-Restriction volumes:
| Part | DNA µl | MQ µl |
|---|---|---|
| SamS II | 12,7 | 13,3 |
| EF III | 8,1 | 17,9 |
| Part | DNA µl | MQ µl |
|---|---|---|
| SamS III | 8,7 | 17,3 |
| EF II | 6,0 | 21,0 |
- Terminator was also digested
Terminator: E/X
| Part | DNA µl | MQ µl |
|---|---|---|
| Terminator | 2,7 | 23,3 |
- Restriction2Test: Agarose gel electrophoresis to validate if the test samples have the correct length. Signals appeared that could not be explained
