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Template:Team:KULeuven/18 August 2009/VanillinProduction
From 2009.igem.org
(Difference between revisions)
(New page: * Cells didn't grow. Perhaps the vector was overloaded. Maybe we can try a Topo-vector? * Run a gel: Restriction products and whole plasmids to compare and check if restriction was succesf...) |
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| - | * Cells didn't grow. | + | * 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'' | ||
Latest revision as of 11:23, 14 September 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
