Team:University of Chicago/Notebook/Gels

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.