Recovery of DNA from Agarose Gel with the Sephaglas BandPrep Kit (Pharmacia)


1. Use TAE buffer for agarose gel electrophoresis.

2. Cut the DNA band which is to be extracted.

3. Cut the slice into several smaller pieces, and transfer to a microfuge tube.

4. For 250mg of the gel slice, add 250 ul of Gel Solubilizer.

5. Vortex vigorously. Incubate the tube for 5-10 min at 60 C.

6. Suspend the Sephaglas BP. Add 5 ul of the suspension to the dissolved gel slice.

7. Vortex gently.

8. Incubate for 5 min at room temperature with occasional agitation.

9. Pulse spin for 30 sec. Remove the supernatant.

10. Repeat the pulse spin for 30 sec. Remove any residual liquid.

11. Add 80 ul of Wash Buffer. Suspend the pellet by pipetting.

12. Pulse spin for 30 sec. Remove the supernatant.

13. Repeat 11 and 12 twice more.

14. Allow the Sephaglas pellet to air-dry for at least 10 min.

15. Add 20 ul of Elution Buffer. Suspend the pellet by pipetting.

16. Incubate for 5 min at room temperature with occasional agitation.

17. Spin at high speed for 1 min. Transfer the supernatant into a new tube.

18. Store the eluted DNA at -20 C until needed.





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