DNA MINI PREP
LYSIS BY BOILING

1. Grow bugs in 0.75 ml LB medium w/ ampicilin O/N @ 37C.
2. Pellet 2-3 min @ 4 K rpm.
3. Remove supernatant (pellet can be stored @ -70C
indefinitelly if needed at this point).
4. Add 100 l STET buffer with
1mg/ml lysozyme and vortex.
5. Boil 2-3 min in heat block (NOT 1 min and NOT 4 min!).
6. Centrifuge 5 min @ 14 K rpm / RT.
7. Remove floppy pellet w/ toothpick.
8. Add 100 l isopropanol and mix by inverting.
9. Store 1 hour @ -20C.
10. Centrifuge 5 min @ 14 K rpm / RT.
11. Pour off the supernatant and wash with 1 ml of 70% EtOH.
12. Spin 2 min @ 14 K rpm.
13. Pour off supernatant and let dry 15-30 min.
14. Resuspend in 40 l H2O.
*** NOTE: Use 10-20 l for restriction enzyme digestion.

Produced and maintained by Peter
Last updated Wed August 9 2000 - 23:06:42