Jonesma Lab:Protocols:miniprep

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  1. Take an overnight E. coli culture and spin down at 3000 rpm for 15min
  2. Discard supernatant and resuspend pellet in 250μL P1 buffer
  3. Add 250μL P2 buffer and mix
  4. Incubate at room temperature for 4min
  5. Add 350μL N3 buffer
  6. Spin at 13 000rpm for 15min
  7. Take supernatant and add to blue miniprep column (with clear plastic reservoir attached)- maximum volume 700μL
  8. Spin at 13 000rpm for 1min
  9. Empty reservoir and add remainder of supernatant from step 7
  10. Spin at 13 000rpm for 1min
  11. Add 700μL PE buffer and incubate for 1min
  12. Spin at 13 000rpm for 1min
  13. Empty reservoir
  14. Spin at 13 000rpm for 1min
  15. Transfer blue column to new 1.5mL tube, labelled appropriately
  16. Add 40μL dH2O and incubate at room temperature for 1min
  17. Spin at 13 000rpm for 1min
  18. Keep tube with DNA, discard used blue column