Gel Extraction

1. Using a razor blade, excise the gel slice containing the DNA fragment of interest under the illumination of LONG wave length UV light (be sure to wear goggles and to cut out as little extra agarose as possible)
2. Measure the weight of the gel slice using a sterile 1.5 ml microcentrifuge tube
3. Add 3 volumes of GEX buffer to 1 volume of gel (100 mg of gel slice is roughly equal to 100 ul)
4. Incubate at 55C on a heating block for 5-10 min until gel is completely dissolved (invert the tube every 1-2 min during incubation)
5. Place an extraction column onto a collection tube. Load 0.7mL dissolved gel mixture onto the column.
6. Centrifuge for 30 seconds at 5000rpm and discard flow through.
7. Repeat step 5 for the remaining mixture if the mixture is more than 0.7mL.
8. Wash the column once with 0.5mL of WN buffer by centrifuging for 30 seconds at 5000 rpm and discard flow through.
9. Wash column once with 0.5mL with WS buffer by centrifuging for 60 seconds at 5000 rpm and discard flowthrough.
10. Centrifuge column at 12000 rpm for another 3 minute to remove residual ethanol.
11. Place column onto new 1.5mL microcentrifuge tube.
12. Add 15-30uL of elution buffer onto the center of the membrane.
13. Stand the column for two minutes and then centrifuge (Remove cap of microcentrifuge tube) for 60 seconds at 12000rpm to elute the DNA.
14. Nanodrop to obtain concentration of DNA.
15. Store at -20 freezer.