Electroduction between Mycobacterium spp. and E. coli

Gregory W. Broussard

Revised: 04/21/10

(Please use, change and disseminate freely!)

1. Pick Mycobacterium colonies carrying your plasmid of choice into 20 µl of ice-cold 10% glycerol.
2. Vortex tube briefly before placing on ice for 10 min.
3. Add Myco-10% glycerol mix to electrocompetent E. coli cells
4. Electroporate using the following parameters: 200 Ω, 2500 V, 25 µF
5. Recover in 900 µl SOC broth at 37 °C for 1 hr.
6. Plate on LB plates with selective antibiotic.
7. Incubate at 37 °C overnight.