Plasmid MaxiPrep Simple

Miniprep

1. Spin down cells 1.5-2 ml of bacterial cultures were pelleted at 6000-7000 rpm for 1 min.
2. Remove culture medium.

3. Add suspension buffer:

5% sucrose, 20–50 mM EDTA, 50 mM Tris pH 8, 0.75 M NH4Cl, 0.5% IGEPAL CA-630 (or Triton X-100), lysozyme 100 µg/ml, and RNase A 25 µg/ml, 20–50 mM CaCl2.

4. Incubated at 65°C for 5 min.
5. Transfer supernatant to another tube.

6. Add 100-120 µl isopropanol,  mix and and spin at 7000 rpm for 10 min at RT.

7. Discard supernatant.

8. Wash with 70% ethanol.

9. Remove ethanol.

10. Dissolve in 50 ul TE.

  100X enzyme stock containing 10 mg/ml lysozyme and 2.5 mg/ml of RNase A prepared in 50% glycerol and 50 mM Tris pH 8 was stored at −20°C and used repeatedly.

Maxiprep

Ref:

http://www.plosone.org/article/info:doi/10.1371/journal.pone.0023457?imageURI=info:doi/10.1371/journal.pone.0023457.t001