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Template:Team:KULeuven/20 August 2009/VanillinProduction
From 2009.igem.org
(Difference between revisions)
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| + | '''1)''' | ||
* EF colonies present! So, we miniprepped them in triple | * EF colonies present! So, we miniprepped them in triple | ||
* Nanodrop concentrations: | * Nanodrop concentrations: | ||
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| + | * We performed a digest with EcoRI and SpeI to cut out the insert and check it's length. Enzymes were added separately, with 1h in between. Total volume was 30µl. | ||
| + | |||
| + | '''2)''' | ||
* Results from yesterday's restriction test were inconclusive, because the DNA ladder didn't run properly so the length of the fragments couldn't be read. Therefore, we did it again... | * Results from yesterday's restriction test were inconclusive, because the DNA ladder didn't run properly so the length of the fragments couldn't be read. Therefore, we did it again... | ||
| - | * Restriction=OK! | + | * Restriction=OK! Test 1 completed. |
| + | |||
| + | |||
| + | '''3)''' | ||
| + | * Test 2: cutting with 2 enzymes (see above) subsequently (1h incubation in between) in a volume of 30µl. Using a larger volume should dilute the glycerol, which inhibits cutting. After that, the products are incubated overnight to allow full restriction. | ||
Revision as of 11:43, 20 August 2009
1)
- EF colonies present! So, we miniprepped them in triple
- Nanodrop concentrations:
| Part | concentration (ng/μl) | 260/280 λ | |
|---|---|---|---|
| EF A | 74,9 | 1,89 | |
| EF B | 63,1 | 1,87 | |
| EF C | 53,9 | 1,90 |
- We performed a digest with EcoRI and SpeI to cut out the insert and check it's length. Enzymes were added separately, with 1h in between. Total volume was 30µl.
2)
- Results from yesterday's restriction test were inconclusive, because the DNA ladder didn't run properly so the length of the fragments couldn't be read. Therefore, we did it again...
- Restriction=OK! Test 1 completed.
3)
- Test 2: cutting with 2 enzymes (see above) subsequently (1h incubation in between) in a volume of 30µl. Using a larger volume should dilute the glycerol, which inhibits cutting. After that, the products are incubated overnight to allow full restriction.
