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Template:Team:KULeuven/18 August 2009/VanillinProduction
From 2009.igem.org
(Difference between revisions)
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* Re-plated the ligation products (SAMS and EF) | * Re-plated the ligation products (SAMS and EF) | ||
| - | * We propose 3 tests to check if the restriction digest (esp. between EcoRI and XbaI, because they lie very close | + | * We propose 3 tests to check if the restriction digest (esp. between EcoRI and XbaI, because they lie very close together) works properly: |
** Cut every plasmid (Sam8, Sam5, Ech, Fcs) separately with EcoRI, XbaI and -only Sam8 and Fcs- with SpeI. | ** Cut every plasmid (Sam8, Sam5, Ech, Fcs) separately with EcoRI, XbaI and -only Sam8 and Fcs- with SpeI. | ||
** Cut subsequently with EcoRI and XbaI, leaving an incubation period of 1h between two digests. Total: 30µl | ** Cut subsequently with EcoRI and XbaI, leaving an incubation period of 1h between two digests. Total: 30µl | ||
** Use pUC18 vector | ** Use pUC18 vector | ||
| - | |||
* Made fluid cultures of Sam8, Sam5, Ech, Fcs | * Made fluid cultures of Sam8, Sam5, Ech, Fcs | ||
Revision as of 13:14, 19 August 2009
- Cells didn't grow. Yet again...
- Re-plated the ligation products (SAMS and EF)
- We propose 3 tests to check if the restriction digest (esp. between EcoRI and XbaI, because they lie very close together) works properly:
- Cut every plasmid (Sam8, Sam5, Ech, Fcs) separately with EcoRI, XbaI and -only Sam8 and Fcs- with SpeI.
- Cut subsequently with EcoRI and XbaI, leaving an incubation period of 1h between two digests. Total: 30µl
- Use pUC18 vector
- Made fluid cultures of Sam8, Sam5, Ech, Fcs
