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Team:KULeuven/13 August 2009
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Latest revision as of 08:47, 10 September 2009
Project progress
Weekly meeting, presentation can be found here
Progress of parts
[edit] Blue Light Receptor
- PCR product of 12-august of pairt
- Miniprep of
Nanodrop results:
| Part | concentration (ng/μl) | 260/280 λ | 260/230 λ |
|---|---|---|---|
| 123,3 | 1,82 | ||
| 141,2 | 1,99 |
- Restriction digest cut with EcoRI and XbaI
- Electroporation of + and the ligation of ...
- Gelelectrophoresis of
- Only 1 band, so probably everything was cut
- Gelextraction of
- Nanodrop = 29,3 ng/μl
- Ligation of vector and insert
[edit] Vanillin Production
- Plated COMT (+RBS) from the registry
- Growing overnight in 37°C
- Plated ech
- From newly made liquid culture
- Miniprep of ech and sam8
- Nanodrop results:
| Part | concentration (ng/μl) | 260/280 λ | 260/230 λ |
|---|---|---|---|
| ech | 72,7 | 1,94 | |
| sam8 | 66,2 | 2,00 |
- Restriction digest on ech and sam8
- Gel electrophoresis on ech and sam8
- New electrocompetent cells were used for the electroporation with the old ligations
- Lig A -> sam5 + sam8
- Lig C -> ech + fcs
- Note: there was no spark
- Plating of the electroporated cells
- 200 μl per plate twice
- 600 μl left of A and C
[edit] Vanillin Receptor
- B and R miniprepped. Tomorrow proceed with mutagenesis of RpoA
- G redone (only TOPO was visible, 200bp) PCR from 2,3 and 1'
- W, X, Y, Z PCR purification to create a TOPO vector tomorrow
Nanodrop:
| Part | concentration (ng/μl) | 260/280 λ | 260/230 λ |
|---|---|---|---|
| W | 46,1 | 1,84 | |
| X | 112 | 1,82 | |
| Y | 207 | 1,85 | |
| Z | 155,3 | 1,86 |
