1. Grow 3 ml overnight culture.
2. Pour in 1.5 ml tube and spin 2' @ 4K RPM.
3. Aspirate supernatant and, add the rest of the bugs
and spin again.
4. Suspend pellet in 200 µl solution I.
5. Add 400 µl solution II, mix by inverting and leave @ RT 10 min.
6. Add 300 µl solution III and leave on ice 10 min.
7. Spin 3 min. @ 14K RPM and remove 800 µl to new tubes.
8. Add 5 µl RNase A (10 mg/ml) and incubate @ 37C 1 hr.
9. Add 250 µl 6 M NaCl, mix and spin 10 min @ 14K RPM.
10. Remove supernatant to two tubes (app. 500 µl in each).
11. Add 450 µl isopropanol to each tube and leave @ RT 10 min.
12. Spin 10 min. @ 14K RPM and wash with 70% EtOH.
13. Resuspend DNA from each tube in 20 µl H₂O and combine.
14. Digest and run 5 µl on agarose gel.
15. From the amount of DNA on the gel determine the amount
needed for sequencing.