QIAprep spin miniprep kit
1. Set up a 5ml overnight culture in selective LB media and incubate at 37oC,
220rpm.
2. Spin down culture at 4000rpm for 10 minutes.
3. Remove supernatant.
4. Resuspend pellet in 250ul Buffer P1 (ensure RNAse has been added) and
transfer to a 1.5ml tube.
5. Add 250ul Buffer P2 and mix thoroughly by inverting the tube 4-6 times (do
not allow reaction to proceed for more than 5 minutes).
6. Add 350ul Buffer N3 and mix immediately by shaking the tube 4-6 times.
7. Centrifuge for 10 min at 13,000 rpm.
8. Apply the resulting supernatant to a labeled QIAprep spin column by
pipetting.
9. Centrifuge for 1 min at 13,000 rpm. Discard the flow-through.
10. Wash the QIAprep spin column by adding 0.5ml Buffer PB and centrifuge for
1 minute at 13,000 rpm. Discard the flow-through.
11. Wash the QIAprep spin column but adding 0.75ml Buffer PE (ensure ethanol
has been added) and centrifuge for 1 minute at 13,000 rpm.
12. Discard the flow-through and centrifuge for an additional 1 minute at 13,000
rpm to remove residual wash buffer.
13. Place the QIAprep spin column in a clean 1.5ml tube. Add 50ul sterile H2O to
the centre of each QIAprep spin column, let it stand for 1 minute and then
centrifuge for 1 minute at 13,000 rpm.