User:Shahar Sukenik/Notebook/p53CT expression in minimal media/2010/08/01

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01/08/10 Transformation

Prep:

1. Prepared LB agar + 1:1000 Amp. plate 2. Pre-heated waterbath to 42C 3. Thawed SOC medium for transformation

Transformation:

1. put 100 ul of C41 cells in snap-cap tube. 2. transfer 1 ul HLT-p53CT plasmid into tube 3. Incubate on ice for 30 min. 4. incubate at 42C for 45 sec. 5. Incubate on ice for 2 min. 6. add 900 ul SOC medium and incubate with shaking at 37C for 1 hour (90 min max) 7. seed 100 ul on LB agar + amp plate using autoclaved glass balls.

Incubation

Incubate O/N at 37C