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Template:Team:KULeuven/14 August 2009/VanillinProduction
From 2009.igem.org
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| - | # Gel extraction of the 4 lanes from | + | # Gel extraction of the 4 lanes from ''sam8'' ({{kulpart|BBa_I742106}}) and ''ech'' ({{kulpart|BBa_I742113 }}) |
| - | #* To test an additional blanc we extracted a blanc piece of gel | + | #* To test an additional blanc, we extracted a blanc piece of gel |
| - | #* Nanodrop results were done with the conventional blanc (just elution buffer) and with the extracted blanco gel. However since the results varied too much | + | #* Nanodrop results were done with the conventional blanc (just elution buffer) and with the extracted blanco gel. However, since the results between both varied too much and the extracted blanco gel was not significantly better, we decided to use the conventional blanco in order to compare between former nanodrop measurements |
Nanodrop: | Nanodrop: | ||
{| border ="1" align="center" | {| border ="1" align="center" | ||
!| Part || concentration (ng/μl) || 260/280 λ || 260/230 λ | !| Part || concentration (ng/μl) || 260/280 λ || 260/230 λ | ||
|- align="center" | |- align="center" | ||
| - | | | + | |''sam8'' ||9,4 || 2,18 || |
|- align="center" | |- align="center" | ||
| - | | | + | | ''ech'' || 13,6|| 1,94 || |
|- | |- | ||
|} | |} | ||
| - | * | + | * Ligation of ''sam5'' (from Wednesday) + ''sam8'' (extracted today) |
{| border ="1" align="center" | {| border ="1" align="center" | ||
!| vector || insert | !| vector || insert | ||
|- align="center" | |- align="center" | ||
| - | | | + | | ''sam5'' || ''sam8'' |
|- align="center" | |- align="center" | ||
| 3637 bp || 1550 bp | | 3637 bp || 1550 bp | ||
| Line 24: | Line 24: | ||
|- | |- | ||
|} | |} | ||
| - | * Ligation of ech and fcs. For fcs we took the digest from last week(C1) , because this | + | * Ligation of ''ech'' and ''fcs''. For ''fcs'' we took the digest from last week (C1), because this week's restriction digest had a very low concentration (4,2) and a very high 260/280 (6,08) |
{| border ="1" align="center" | {| border ="1" align="center" | ||
!| vector || insert | !| vector || insert | ||
|- align="center" | |- align="center" | ||
| - | | ech || fcs | + | | ''ech'' || ''fcs'' |
|- align="center" | |- align="center" | ||
| 2929 bp || 1789 bp | | 2929 bp || 1789 bp | ||
| Line 37: | Line 37: | ||
|- | |- | ||
|} | |} | ||
| - | * Cells were stored | + | * Cells were stored at 16°C |
* Plated the electroporated cells from the A and C ligation | * Plated the electroporated cells from the A and C ligation | ||
| - | * | + | * Made Agar and poured new plates with Ap |
Latest revision as of 10:04, 14 September 2009
- Gel extraction of the 4 lanes from sam8 () and ech ()
- To test an additional blanc, we extracted a blanc piece of gel
- Nanodrop results were done with the conventional blanc (just elution buffer) and with the extracted blanco gel. However, since the results between both varied too much and the extracted blanco gel was not significantly better, we decided to use the conventional blanco in order to compare between former nanodrop measurements
Nanodrop:
| Part | concentration (ng/μl) | 260/280 λ | 260/230 λ |
|---|---|---|---|
| sam8 | 9,4 | 2,18 | |
| ech | 13,6 | 1,94 |
- Ligation of sam5 (from Wednesday) + sam8 (extracted today)
| vector | insert |
|---|---|
| sam5 | sam8 |
| 3637 bp | 1550 bp |
| 50 ng | 85,2 ng |
| 7 μl | 9 μl |
- Ligation of ech and fcs. For fcs we took the digest from last week (C1), because this week's restriction digest had a very low concentration (4,2) and a very high 260/280 (6,08)
| vector | insert |
|---|---|
| ech | fcs |
| 2929 bp | 1789 bp |
| 50 ng | 122 ng |
| 3 μl | 9 μl |
- Cells were stored at 16°C
- Plated the electroporated cells from the A and C ligation
- Made Agar and poured new plates with Ap
