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The magic E. coli transformation protocol

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The magic E. coli transformation protocol

 

  1. Take the ultra-competent cells out of freezer. Thaw on ice.
  2. Put your selection plates (LB +?) on the bottom shelf of an incubator to pre-warm them (about 37 to 45o C)
  3. Add half of your ligation to the cells, leave on ice for 5’.
  4. Add the cell/ligation mix onto the pre-warmed plates (2-3 plates: 10ul; 50ul; 200ul), turn the plate quickly to spread the cells.
  5. Spread the cells evenly onto the plates. Airdry the plates in a hood for 5 min. and culture o/n.

 

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