Editing Extaction of DNA from Agarose Gel

1. Run DNA on a 1% TAE agarose gel&nbsp;<br/>2. Excise the gel slice with gel cutter<br/>3. Place gel in eppenorff tube.<br/>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.<br/>6. Ensure that the gel has completely dissolved.<br/>7. Pipette 750μl into one of the columns.<br/>8. Centrifugethe column for a few secs (typically up to top speed and then stop is sufficient).<br/>9. Discard the flow through and add the remaining sample to the column.<br/>10. Repeat#7.10. Discardthe flow through.<br/>11. Add 750μl of PE buffer (with alcohol added as per instructions on bottle) to the column&nbsp;

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