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Template:Team:KULeuven/26 August 2009/VanillinProduction
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Revision as of 12:44, 27 August 2009 by JochemDeen (Talk | contribs)
- 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 | 17,3 |
| EF III | 8,1 | 21,9 |
| Part | DNA µl | MQ µl |
|---|---|---|
| SamS III | 8,7 | 21,3 |
| EF II | 6,0 | 24,0 |
- Terminator was also digested
Terminator: E/X
| Part | DNA µl | MQ µl |
|---|---|---|
| Terminator | 2,7 | 27,3 |
- Agarose gel electrophorese to validate if the test samples contain the correct length.
