Purify DNA from Gel using Isopropanol

1, Make <1% agarose gel.

2, Run DNA on gel, cut out band into a microcentrifuge tube

3. Add  1~3 vol of QG Buffer. 

4. Use warm water bath (60~90 oC) to dissolve gel.

5. Measure final vol using pipette. 

6. Add 0.6 X isopropanol. Mix

7. Spin at max speed for 5 min.

8. Dump off liquid.

9. Add 100 ul 70% ethanol. 

10. Spin 1 min.

11. Dump off ethanol.

12. Spin 5 seconds.

13. Remove liquid using 10 ul pippett.

14. Dry at 50 oC on heat block for 1 min.

15. Add 5~20 ul ddH2O. Votex.

16.  Continue to digestion or ligation. 

Lane 1: Recover DNA from gel by centrifugation for 15 min using a spin column, then precipitate using isopropanol

Lane 2: Recover DNA from gel by dissolving gel in QG buffer then then precipitate DNA with 0.6 X isopropanol