Mini Preps

from a 2 ml culture of bacteria in TB grown overnight

1. Pour 1.5 ml into an eppendorf tube.

2. Centrifuge at 3 000 rpm for 3 min. at room temperature.

3. Discard supernatant.

4. Redisolve pellet in 100ul solution 1.

5. Vortex

6. Add 200 ul solution 2

7. Mix thoroughly by inversion.

8. Put on ice for 5 min.

9. Add 150 ul solution 3

10. Mix thoroughly by inversion.

11. Put on ice for 5 min

12. centrifuge at max speed for 10 min. at 4C.

13. Take off 360 ul supernatant.

14. Transfer into an eppendorf tube with 800 ul 100% EtOH.

15. Store for 5 min. at room temperature.

16. Precipitate by centrifuging at max speed for 15 min. at 4C.

17. Wash pellet with 70% EtOH.

18. Airdry pellet.

19. Redisolve in 20ul TE containing RNAse.


Solution 1: 50 mM glucose, 10 mM EDTA pH8.0, 25 mM Tris pH7.0, autoclave at 100C.

Solution 2: 2% SDS + 0.4 N NaOH , mix these two just before use.

Solution 3: 5M Potassium Acetate.




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