1. Grow O/N in 1.5 ml LMM or Terrific broth (see Reagents) with 75µg/ml Amp
2. Pour into ependorf tube and spin down cells at 7-8K for 2 min
3. Aspirate s/n and resuspend in 50µl 25mM Tris pH 8, 10 mM EDTA; leave lids open
4. Add 100µl of freshly prepared 1% SDS, 0.2M NaOH (5ml = 100ul 10M NaOH added to 4.4ml DDW then 500µl 10% SDS). Add it forcefully and you don't need to vortex
5. Add 75µl KoAc solution and vortex
6. Add 100µl of phenol/CHI₃, close lids, vortex
7. Spin 13K for 2 mins
8. Remove supernatant, add to 500µl ethanol. Vortex and spin at 13 K for 5 min
9. Aspirate s/n, removing all ethanol
10. Resuspend in 50µl TE
11. Digest 2-5ul, adding 1µl preboiled 10mg/ml RNAse A (see Reagents for preparation).

KOAc solution:

NOTE: Large scale plasmid preps can be made quickly by scaling all reagents of the "mini prep" procedure 100-fold. Once the phenol step has been completed treat the supernatant with 50ul 10mg/ml RNA'se for 1hr, isopropanol ppte (0.6volumes) and resuspend DNA in 400µl of TE. reprecipitate. If the ppte is very cloudy (residual RNA) hook the DNA out (fluffy ball) to separate it from RNA.