Team:DTU-Denmark/Methods/Plasmid isolation

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Plasmid Isolation

  1. Follow QIAprep Spin Miniprep Kit protocol from steps 1 - 4.
  2. Centrifuge for 1h at 4C and 20000 g.
  3. Carefully transfer the supernatant to a new Eppendorf. Be careful not to take any solid parts (this is the cell debris).
  4. To the sample add 3 volumes of ice-cold 70% ethanol. If needed divide into several samples.
  5. Put samples to -20 for 0.5h to 1 h to precipitate DNA.
  6. Centrifuge at 4C for 30 mins at 20000 g.
  7. Remove the supernatant.
  8. Add 300 uL of 90% ice-cold ethanol. Do not pipette up and down.
  9. If the pellet is detached centrifuge for 5 to 10 mins at 4C and 20000 g.
  10. Remove the ethanol with a pipette.
  11. Let the sample stand open to dry for up to 5 mins.
  12. Resuspend in 50 - 100 uL water.

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