Plasmid miniprep --- boiling

Edited by Chang Zhu
1. Transfer 1 ml of bacteria culture to microfuge tube. Centrifuge 1 minute at maximum speed to pellet cells. Discard supernatant.

2. Add 100 ml STET buffer containing lysozyme (8% sucrose, 0.5% TRITON X-100, 50 mM EDTA, 10 mM TRIS-Cl pH 8.0 and 1 mg/ml lyzozyme. Add lysozyme fresh.)

3. Vortexing.

4. Place in boiling water bath for 40 seconds.

5. Put the tube immediately on ice for a few minutes.

6. Centrifuge 10 minute at maximum speed.

7. Transfer the supernatant to another tube or using a toothpick, pick pellet out and discard.

8. Add 100 ml of isopropanol to supernatant, mix.

9. Centrifuge 10 minute at maximum speed. Discard supernatant.

10. Wash pellet with 70% ethanol.

11. Air dry.

12. Resuspend DNA in 20-50 ml of TE or TE/10 mg /ml RNAse A.