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1. Pellet 1-5mL of bacterial overnight culture by centrifugation at >8000 rpm for 3 min at room temperature (15-25C; use 2ml microcentrifuge collection tubes). Decant all the liquid and add 1 ml of the culture into the corresponding tube. Make sure not to mix up the tries.
2. Resuspend pelleted bacterial cells in 250 uL Buffer P1 and transfer to microcentrifuge tube.
3. Add 250uL Buffer P2 and mix thoroughly by inverting tube 4-6 times. Do NOT vortex. Mixture turns blue. Do NOT allow this lysis reaction to proceed for more than 5 min.
4. Add 350uL of Buffer N3 and mix IMMEDIATELY and thoroughly by inverting tube 4-6 times. Do NOT vortex. Mixture is now colorless.
5. Centrifuge for 10min at 13,000 rpm in table-top centrifuge.
6. Apply the supernatant to a QIAprep spin column by decanting or pipetting. Do NOT get any of the sticky precipitate.
7. Centrifuge for 30 - 60s at 13000rpm. Discard flow-through.
8. Wash the QIAprep column by adding 0.5 mL Buffer PB.
9. Centrifuge for 30 - 60s at 13000rpm. Discard flow-through.
10. Wash the QIAprep column by adding 0.75 mL Buffer PE.
11. Centrifuge for 30 - 60s at 13000rpm. Discard flow-through.
12. Centrifuge for 1 min to remove residual was buffer.
13. Place the QIAprep column in a clean 1.5mL microcentrifuge tube. To elute DNA, add 50uL Buffer EB to center of each column. Be careful NOT to pierce column.
14. Let stand for 1 minute.
15. Centrifuge for 60s at 13000rpm.
16. Remove column and discard, tube now contains DNA.
17. Go to NanoDrop and spec DNA.

http://2014.igem.org/wiki/images/f/f5/QIAprep-Spin-Miniprep-Kit-EN.pdf