Team:University of Chicago/Notebook/Gels

From 2008.igem.org

DNA and protein gels

Agar

0.8%

1. Weigh out .8g agarose
2. Put agarose in 500mL Erlenmeyer flask.
3. Add 100mL 1X TBE Buffer. Swirl.
4. Microwave at 100% power until the agarose starts to dissolve. Every 30-40 seconds, stop and swirl. Continue microwaving until solution boils and agarose is dissolved.
5. After agarose is dissolved, cover flask with foil or aran wrap and place in 55-60C waterbath until needed.
6. When Agarose has cooled to enough to touch comfortably, add 10microliters Syber Safe. Swirl.
7. Pour 30mL gels. Use 50Ml conical tube to measure 30mL.

1.2%

Same as above but use 1.2 g agarose instead of .8g

Coomassie Blue Stain/Destain Recipe

450 ml water
450 ml methanol
100 ml 100% (glacial) acetic acid

1. Add 2 g per liter coomassie blue for stain. Destain is exactly the same, except do not add coomassie blue.
2. To stain the gel, incubate it with gentle shaking for at least 30 minutes. Pour the stain back into the bottle (you can reuse it). Rinse out the residual stain with a small amount of destain solution, and then incubate with gentle shaking for ~1 hour or more. To speed up destaining, you can put some folded KimWipes in the dish. The dye will adsorb to these, which will remove it from solution.
3. After your gel is destained, you can rehydrate it in distilled water and then image it and/or place it on a piece of filter paper and dry it with a gel dryer.

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