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Template:Team:KULeuven/18 August 2009/VanillinProduction
From 2009.igem.org
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* We propose 3 tests to check if the restriction digest (esp. between EcoRI and XbaI, because they lie very close together) works properly: | * We propose 3 tests to check if the restriction digest (esp. between EcoRI and XbaI, because they lie very close together) works properly: | ||
| - | + | '''Test 1''': Cut every plasmid (Sam8, Sam5, Ech, Fcs) separately with EcoRI, XbaI and -only Sam8 and Fcs- with SpeI. | |
| - | + | ||
| - | + | '''Test 2''': Cut subsequently with EcoRI and XbaI, leaving an incubation period of 1h between two digests. Total volume: 30µl, to dilute glycerol (which inhibits restriction) | |
| + | |||
| + | '''Test 3''': Use pUC18 vector | ||
* Made fluid cultures of Sam8, Sam5, Ech, Fcs | * Made fluid cultures of Sam8, Sam5, Ech, Fcs | ||
Revision as of 11:59, 20 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:
Test 1: Cut every plasmid (Sam8, Sam5, Ech, Fcs) separately with EcoRI, XbaI and -only Sam8 and Fcs- with SpeI.
Test 2: Cut subsequently with EcoRI and XbaI, leaving an incubation period of 1h between two digests. Total volume: 30µl, to dilute glycerol (which inhibits restriction)
Test 3: Use pUC18 vector
- Made fluid cultures of Sam8, Sam5, Ech, Fcs
