Template:Team:KULeuven/29 July 2009/BlueLightReceptor

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**:the mixture was incubated for 15 min at 37°C
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**the mixture was incubated for 15 min at 37°C

Revision as of 21:07, 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:
I II III IV V VI
EcoRI digestion mix 20μl 20μl 20μl 20μl 20μl 20μl
diluted SpeI 1μl 1/200 2μl 1/200 3μl 1/200 1μl 1/500 2μl 1/500 3μl 1/500
AD 4μl 3μl 2μl 4μl 3μl 2μl
    • the mixture was incubated for 15 min at 37°C
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