KCM Competent Transformation Protocol



KCM Competent Transformation Protocol

1. Thaw an aliquot of KCM competent cells on ice.

2. Remove 100 uL from the aliquot and place it in a fresh tube.

3. Add 30 uL 5x KCM solution and 50 uL ddH20 to the cells.

4. Add your DNA (1 uL of prepped plasmid is more than sufficient).

5. Incubate cells on ice for 5 minutes.

6. Heat shock cells at 42 C for 1-2 minutes.

7. Add 100 uL prewarmed SOC and allow cells to recover at 37 C for 1 hour.

5x KCM Solution

0.5 M KCl

0.15 M CaCl2

0.25 M MgCl2

Make 10 mL of 5x KCM solution, filter sterilize it, and store it at 4 C.

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