Editing Plasmid MaxiPrep HIC (section)

== For 500 ml ~ 1 L culture ==

1. Grow high copy number plasmid in DH5alpha in 1 L LB/Amp or LB/Kan o/n at 37oC. 2. Spin down cells

3. Resuspend cells with 20 ml P1 buffer (no RNase).

4. Lyse cells with 20 ml of P2 for 5 min.

5. Neutralize with 20 ml of P3.

6. Cool on ice for 5 min.

7. Spin at 6000 rpm for 10 min, high speed centrifuge, collect supernatant, filter liquid using coffee filter if necessary.<br/>

E. Coli genomic DNA and RNA were precipitated by addition of solid ammonium sulfate at 80% by weight saturation. The pH of the solution was maintained at 5.2+/−0.2. The suspension was allowed to stir slowly overnight at 4° C. The resulting precipitate (containing the genomic DNA and RNA) was; removed by centrifugation at 10,000× g for 30 min.