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Template:Team:KULeuven/20 August 2009/VanillinProduction
From 2009.igem.org
(Difference between revisions)
| Line 13: | Line 13: | ||
* 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. | * 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. | ||
| + | * {| border ="1" align="center" | ||
| + | !| Part || µl DNA || µl MQ || | ||
| + | |- align="center" | ||
| + | | EF C ||9,3 || 20,7 || | ||
| + | |- | ||
| + | |} | ||
'''2)''' | '''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... | + | * [[Test 1]]: 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. | * Restriction=OK! Test 1 completed. | ||
'''3)''' | '''3)''' | ||
| - | * Test 2: cutting with 2 enzymes | + | * [[Test 2]]: cutting with 2 enzymes 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. Sam8 and Fcs are cut with EcoRI and SpeI; Sam5 and Ech are cut with EcoRI and XbaI |
| + | |||
| + | *{| border ="1" align="center" | ||
| + | !| Part || µl DNA || µl MQ || | ||
| + | |- align="center" | ||
| + | | Sam8 B ||7,9 || 22,1 || | ||
| + | |- align="center" | ||
| + | | Sam5 B ||4,0 || 26,0 || | ||
| + | |- align="center" | ||
| + | | Ech B ||5,2 || 24,8 || | ||
| + | |- align="center" | ||
| + | | Fcs B ||4,4 || 25,6 || | ||
| + | |- | ||
| + | |} | ||
Revision as of 11:54, 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.
- {| border ="1" align="center"
!| Part || µl DNA || µl MQ || |- align="center" | EF C ||9,3 || 20,7 || |- |}
2)
- Test 1: 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 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. Sam8 and Fcs are cut with EcoRI and SpeI; Sam5 and Ech are cut with EcoRI and XbaI
- {| border ="1" align="center"
!| Part || µl DNA || µl MQ || |- align="center" | Sam8 B ||7,9 || 22,1 || |- align="center" | Sam5 B ||4,0 || 26,0 || |- align="center" | Ech B ||5,2 || 24,8 || |- align="center" | Fcs B ||4,4 || 25,6 || |- |}
