Template:Team:KULeuven/29 July 2009/BlueLightReceptor
From 2009.igem.org
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 | |
---|---|---|---|---|---|---|
x | x1 | x2 | x3 | x4 | x5 | x6 |
y | y1 | y2 | y3 | y4 | y5 | y6 |
z | z1 | z2 | z3 | z4 | z5 | z6 |