05/09/13

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Transformation of pSB1C3/TOD genes

  1. Centrifuge the ligation
  2. Add 6 ul of each ligation reaction to a sterile 1.5ml tube on ice
  3. Thaw the cells on ice
  4. Carefully transfer 50ul of cells to the ligation reaction. Gently flick the tubes and incubate on ice for 20 minutes.
  5. Heat shock the cells for 45 seconds in a 42'C water bath. Do not shake. Immediately return the tubes to ice for 2 minutes.
  6. Add 950 ul of SOC medium (at room temperature) to the ligation reactions. Incubate for 1.5 hours at 37'C on a shaker. 
*The controls for this experiment are the resistance and viability controls. 
*One tube has 50 ul of highly competent cells and SOC medium added to it.
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