PCR Template from M. smegmatis Culture

Gregory W. Broussard

Revised:  9/10/10

(Please use, change and disseminate freely!)

1. Place 1 mL culture in a 1.5 mL tube
2. Centrifuge at 5,000 X g for 5 min.
3. Resuspend pellet in 500 µL TE
4. Place at 95°C for 20 min.
5. Place on ice for ~1 min. VORTEX on high setting!!!
6. Centrifuge to pellet, 13,000 rpm for 1 min.
7. Save supernatant in a 1.5 mL tube
8. Use 10 µL of this as the template in PCR reactions