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