Team:Osaka/week5

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)
7. Dilution of primer

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