Template:Team:KULeuven/29 July 2009/BlueLightReceptor

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(Difference between revisions)
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**a mixture of 20μl was made:6μl DNA, 2μl bufferH, 1μl EcoRI and 1μl XbaI, 10μl AD
**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
**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:

Revision as of 12:05, 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:
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