Team:KULeuven/7 September 2009
From 2009.igem.org
Project progress
Progress of parts
[edit] Blue Light Receptor
- Restriction digest with EcoRI and PstI on and the purified pcr product of in order to get our promotor on itself in the standard vector.
- Gel electroforesis and extraction of this RD. The concentration of this extraction was too low so no ligation was performed with these samples.
- Extra pcr was done on with primers 2260 and 2261.
- Liquid cultures of LigX and of were made from the 4x plates.
[edit] Vanillin Production
- Miniprep of the four colonies from EF ligation
- Accidentally used double volumes of elution buffer on EF4
part | concentration | 260/280 | 260/230 |
---|---|---|---|
EF 1 | 115,5 | 1,98 | 2,31 |
EF 2 | 105,2 | 1,96 | 1,66 |
EF 3 | 95,1 | 2,05 | 0,98 |
EF 4 | 52,7 | 2,06 | 0,97 |
- Gel electrophoresis from restriction on sam5 and sam8 looked good
- Purified and ligated the restriction from sam5 and sam8 overnight
- Concentration of sam8 was barely enough for ligation...
part | concentration | 260/280 | 260/230 |
---|---|---|---|
sam5 | 10,1 | 2,11 | 0,04 |
sam8 | 3,1 | 3,96 | 0,02 |
[edit] Vanillin Receptor
- a new restrictiondigest was done for puc and A because we decided that earlier results were not good enough.
- There was a gelextraction, nanodrop and ligation done overnight.
- The colony PCR of rpoA => 428 again for one minute => bad result
- Miniprep of X+K was checked and put on gel. Wrong result probably because we selected a wrong colony.