Huang: DH5alpha transformation

Overview

This protocol allows competent cell transformation.

Materials

• DH5alpha competent cells
• LB
• LB + antibiotic plates
• Plasmid DNA or DNA to transform the cells with

Procedure

• 1. Set tubes of frozen cells on ice and let thaw very slowly
• 2. Add 100ul thawed cells to new eppendorf
• 3. Add 1-2ul DNA to cells
• 4. Incubate on ice for 20 min
• 5. Heat shock in 42C water bath for one min (make sure water bath is at exactly 42C)
• 6. Incubate on ice 2 min
• 7. Add 900ul growth medium (LB) and shake 1 hour at 37C
• 8. Plate about 200ul and grow overnight

Contact

• Carolina Tropini