Extaction of DNA from Agarose Gel

1. Run DNA on a 1% TAE agarose gel 
2. Excise the gel slice with gel cutter
3. Place gel in eppenorff tube.
4. Add QG buffer to the top of the tube–add at least 1 ml; if it does not look like 1 ml willfit, cut the gel slice into 2 pieces and put into 2 tubes and process both tubes as individual samples.

5. Incubatethe gel slice in QG buffer at 50 oC running water for approx 10 minutes, mix every couple of minutes.
6. Ensure that the gel has completely dissolved.
7. Pipette 750μl into one of the columns.
8. Centrifugethe column for a few secs (typically up to top speed and then stop is sufficient).
9. Discard the flow through and add the remaining sample to the column.
10. Repeat#7.10. Discardthe flow through.
11. Add 750μl of PE buffer (with alcohol added as per instructions on bottle) to the column 

12. Centrifugethe column for a few secs (typically up to top speed and then stop is sufficient).
13. Discardthe flow through.
14. Centrifugethe column for 1 minute at top speed (this ensures that all the alcohol is removed from the column).
15. Placethe column in a fresh tube.
16. Add 15 μl EB (or use dH20 directly onto the disc at the base of the column (do not touch the disc).
17. Stand at RT for 1 minutes.
18. Centrifuge at top speed for 1 minute.
19. Discard column.
20. Put eluted DNA on ice.
21. Continue to ligation or store eluted DNA at -20 oC (-20 oC freezer).