Team:UT Dallas/NotebookWeek6

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biz solution

Week 6

August 15-

  • PCR of ptetR, RBS, LuxI, term
  • Gel electrophoresis and purification
  • Glycerol stock and miniprep
  • Incubation in 3 mL LB can overnight at 37 degrees Celsius and 220 rpm
  • Test digestion for 1 hour and 30 minutes at 37 degrees Celsius
  • Gel Electrophoresis
  • Test Digestion Observation: Lane 5 gave positive results for FGF-R: 2 bands observed at predicted bands size. Lane 4, 7, 9 were all negative bad results because EcoRI was used but only 1 band showed. This suggests that FGFR was not inserted. Lanes 10, 11, 12 were CheZ* and all matched negative control. Results are bad for CheZ*- will be regrown/miniprep/ligated.
  • Incubation in 3 mL LB chlora overnight at 37 degrees Celsius and 220 rpm
  • August 16-

  • Digestion; Gel Electrophoresis and Purification
  • Ligation (10 minutes at room temperature)
  • Transformation (50 µL DH5α, 2 µL DNA, 37 degrees Celsius overnight, all carb)
  • Re- incubating – taking 20 µL of yesterday’s incubations and placing them in 3 mL of LB and appropriate
  • Transformation (50 µL DH5α, 2 µL DNA, 37 degrees Celsius overnight, all carb)
  • August 17-

  • Glycerol stock ToxR and CheZ* colonies (2nd attempt)
  • Miniprep
  • no results on gel
  • Incubate in 3 mL LB carb overnight at 37 degrees Celsius and 220 rpm
  • Transformation (50 µL DH5α, 2 µL DNA, 37 degrees Celsius overnight, all carb)
  • Incubation in 3 mL of LB chlora overnight at 37 degrees Celsius and 220 rpm
  • August 18-

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    Notebook

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