Team:Osaka/week5

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August 26 (Sun)

  1. Colony check
    • 1-1G,1-5A → Successfully transformed
    • 5-23O,1-7A → Transformed cells didn't grow on plates
  2. Transfer to liquid culture:1-1G,1-5A
  3. Preparation of LB agar plates (Tet)

August 27 (Mon)

  1. miniprep(1-1G,1-5A)
  2. Transformaton of ligation products into DH5α E.coli:155,156
    • Transformed cells did not grow on plates

August 28 (Tue)

  1. Preparation of LB agar plates (Amp,Cam)
  2. Ligation(155,156)
  3. Transformaton of ligation products into DH5α E.coli:155,156,152(positive control)
  4. To liquid culture(152,155,Rosetta wild type)

August 29 (Wed)

  1. miniprep(155①~④)
  2. Restriction digests(155,1-1G)
  3. Electrophoresis(153,1-1G)
  4. Transformation(156)
  5. To liquid culture(152,155,WT)
    • 155 → turned red

August 30 (Thu)

  1. Preparation of LB agar plates (Amp,Cam)
  2. Electrophoresis(1-1G,1-5A,1-7A,psBIC3)
    • 1-5A,1-7A → 3 bands appered on a gel (contaminated)
  3. Tolerance assay(preliminary test,MitomycinC)
  4. Gel extraction(1-1G,1-5A,1-7A,1-3A)
  5. Ligation(155,156)
  6. Transformation(155,156)
    • 155 → Successfully transformed
    • 156 → Transformed cells did not grow on plates

August 31 (Fri)

  1. Colony check
  2. Preparation of LB agar plate
  3. Tolerance assay (preliminary test,MitomycinC)

September 1 (Sat)

  1. Miniprep
  2. Tolerance assay(preliminary test2,MitomycinC)
  3. Restriction digests(155①,②)
  4. Electrophoresis
  5. Transformation(156)
  6. To liquid culture(152,155)


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