Team:UNICAMP-EMSE Brazil/protocols/Transforming bacteria

Transforming bacteria:

Thermocompetent Transformation

• In the flux, add between 5 and 10 μl of ligation or purified pDNA to 15 μl of transformation buffer.
• Add the mixture to the tube containing competent bacteria and pipette up and down.
• Place the tube on ice for 30 min.
• Leave the tube for 10 min in room temperature.
• Add 1ml of LB-Media

• 1 hour at 37˚C.

Plate aliquotes of the transformed bacteria on LB-agar containing the appropriate antibiotic.

Incubate overnight at 37˚C.

See how to make competent cells