Purification of Plasmid by the FlexiPrep Kit (Pharmacia)


1. Transfer 1.5 ml of an overnight culture of E. coli to a microfuge tube.

2. Collect cells by spinning for 1 min.

3. Resuspend the cell pellet in 200 ul of Solution I.

4. Add 200 ul of Solution II.

5. Add 200 ul of Solution III.

6. Spin for 5 min at room temperature

7. Transfer the supernatant into a new microfuge tube.

8. Add 420 ul of isopropanol. Spin for 10 min. Discard the supernatant.

9. Add 150 ul of the suspension of Sephaglas™ FP slurry.

10. Vortex gently for 1 min to dissolve the pellet.

11. Spin for 15 sec. Discard the supernatant.

12. Add 200 ul of Wash Buffer. Vortex gently.

13. Spin for 15 sec. Discard the supernatant.

14. Add 300 ul of 70% ethanol. Vortex gently.

15. Spin for 15 sec. Discard the supernatant.

16. Allow the pellet to air dry for 10 min.

17. Add 50 ul of TE. Incubate for 5 min at room temperature.

18. Spin for 1 min. Transfer the supernatant to a new microfuge tube.





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