Promega Wizard Plus SV Mini-Prep

1. For each miniprep, grow 2 ml culture, 37° C, overnight (O/N) in appropriate medium and anitbiotic (usually ampicillin,but not always); shake at 400 RPM and slant tubes.
   
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2. Pour the contents O/N culture into one labeled tube. Replace the metal cap and save the culture tube at 4° C.
3. Spin the microfuge tube for 2 min.
4. Aspirate off the medium.
5. Resuspend pellet in 250 µl Cell Resuspension Solution. Resuspend cells very well by pipetting up and down.
6. Add 250 µl of Cell Lysis Solution. Mix by inverting the tube 4 times.
7. Add 10 µl Alkaline Protease Solution. Mix by inverting the tube 4 times. Incubate 3 minutes at room temperature (RT°).
8. Add 350µl Neutralization Solution. Mix by inverting the tube 4 times.
9. Spin full speed for 10 minutes at RT°.
10. Prepare Spin Column (with binding resin) by inserting into 2 mL collection tube.
11. Transfer supernatant to Spin Column.
12. Spin full speed for 1 minute at RT°. Discard liquid flowthrough and reinsert Spin Column to collection tube.
13. Add 750 µl Wash Solution (with ethonal already added).
14. Spin full speed for 1 minute at RT°. Discard liquid flowthrough and reinsert Spin Column to collection tube.
15. Add 250 µl Wash Solution (with ethonol already added).
16. Spin full speed for 1 minute at RT°. Discard liquid flowthrough and transfer Spin Column to a clean 1.5 mL microfuge tube.
    Be sure to label the spin column and the microfuge tube.
17. Add 100 µl Nuclease-Free Water to the Spin Column.
18. Spin full speed for 1 minute at RT°. SAVE THE LIQUID with your plasmid. Discard the spin column. Store DNA at -20° C.
19. If you want to digest some DNA, 5-10 µl of the MP DNA in a final volume of 20 µl is a nice place to start.