Heat-Shock Transformation

Reagents:

1. 50ul of chemically competent cells

2. 22,5ul of MgCl2

3. 1ul plasmid

Steps:

1. Allow chemically competent cell thawed for 1 hour on ice.

2. Add to the cells 22,5ul of MgCl2 and 1ul of plasmid, leaving on ice for 30 minutes.

3. Submit a heat-shock of 42 °C for 45 seconds.

4. Incubate the cells on ice for 10 minutes.

5. Add 500ul of LB medium and incubated in the shaker at 37 ° C for 50 to 60minutes.

6. Subculturing in LB agar with antibiotic selection.

Do not forget the negative control!