Template:Team:KULeuven/29 July 2009/BlueLightReceptor
From 2009.igem.org
(Difference between revisions)
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! class="top" | VI | ! class="top" | VI | ||
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- | ! class="left" | | + | ! class="left" | EcoRI digestion mix |
| x1 ||x2 || x3 || x4 || x5 || x6 | | x1 ||x2 || x3 || x4 || x5 || x6 | ||
|- | |- | ||
- | ! class="left" | | + | ! class="left" | diluted SpeI |
| y1 || y2 || y3 || y4 || y5 || y6 | | y1 || y2 || y3 || y4 || y5 || y6 | ||
|- | |- | ||
- | ! class="left" | | + | ! class="left" | AD |
| z1 || z2 || z3 || z4 || z5 || z6 | | z1 || z2 || z3 || z4 || z5 || z6 | ||
|- | |- | ||
|} | |} |
Revision as of 13:37, 29 July 2009
GFP ()
- miniprepped and nanodropped
- concentration: 85,6ng/μl
- 260/280: 1,87
- a restriction digest was performed to cut the plasmid with EcoRI and XbaI
- a mixture of 20μl was made:6μl DNA, 2μl bufferH, 1μl EcoRI and 1μl XbaI, 10μl AD
- the mixture was incubated for at least an hour at 37°C
BLR promoter region The PCR product that was purified friday (24/07) is digested with EcoRI and partially digested with SpeI
- digestion with EcoRI
- following mixture was made (x6): 5μl DNA, 2μl bufferH, 1μl EcoRI, 12μl MilliQ
- incubated for 1h at 37°C
- partial digestion with SpeI
- dilution of the enzymes:
- AD/b: 225μl MQ + 25μl bufferH
- 1/100: 1μl SpeI + 99μl AD/b
- 1/200: 50μl 1/100 + 50μl AD/b
- 1/500: 20μl 1/100 + 80μl AD/b
- making following mixture:
- dilution of the enzymes:
I | II | III | IV | V | VI | |
---|---|---|---|---|---|---|
EcoRI digestion mix | x1 | x2 | x3 | x4 | x5 | x6 |
diluted SpeI | y1 | y2 | y3 | y4 | y5 | y6 |
AD | z1 | z2 | z3 | z4 | z5 | z6 |