Promega Maxi Prep (without vacuum manifold)

  1. Pellet 100-500ml cell solution in a centrifuge bottle at 4,986 rpm (5,000g) for 10 min.
  2. Discard supernatant, Resuspend in 15ml Cell Resuspension Solution
  3. Add 15ml Cell Lysis Solution, invert up to 20 times
  4. Add 15ml Neutralization Solution, invert several times
  5. Spin at 8,344rpm (14,000g) for 15min
  6. Run supernatant through filter paper into a graduated cylinder
  7. Transfer to a new centrifuge bottle and add 50% volume isopropanol
  8. Spin at 8,344rpm (14,000g) for 15min
  9. Discard supernatant, resuspend in 2ml TE
  10. Add 10ml DNA Purification Resin, swirl
  11. Pour into shredder column, let drain
  12. Spin at 2,500rpm (1,300g) with a swinging basket rotor for 5min, pour off
  13. Add 25ml Column Wash Solution
  14. Spin at 2,500rpm (1,300g) with a swinging basket rotor for 5min
  15. Add 5ml 80% ethanol, let drain
  16. Spin at 2,500rpm (1,300g) with a swinging basket rotor for 5min, pour off
  17. Spin again, discard collection tube
  18. Place column in a new 50ml tube and add 1.5ml 65° water, wait 1 min
  19. Spin at 2,500rpm (1,300g) with a swinging basket rotor for 5min
  20.  Pour into syringe/filter set up and filter into a 1.5ml tube
  21. Spin at 8,344rpm (14,000g) for 1 min
  22. Transfer supernatant to a new tube.