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Team:University of Chicago/Notebook/Gels
From 2008.igem.org
(Difference between revisions)
(New page: === DNA and protein gels === ==== Agar ==== ===== 0.8% ===== # Weigh out .8g agarose # Put agarose in 500mL Erlenmeyer flask. # Add 100mL 1X TBE Buffer. Swirl. # Microwave at 100% power un...) |
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===== 1.2% ===== | ===== 1.2% ===== | ||
Same as above but use 1.2 g agarose instead of .8g | Same as above but use 1.2 g agarose instead of .8g | ||
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====Coomassie Blue Stain/Destain Recipe==== | ====Coomassie Blue Stain/Destain Recipe==== | ||
Latest revision as of 18:43, 29 July 2008
Contents |
DNA and protein gels
Agar
0.8%
- Weigh out .8g agarose
- Put agarose in 500mL Erlenmeyer flask.
- Add 100mL 1X TBE Buffer. Swirl.
- 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.
- After agarose is dissolved, cover flask with foil or aran wrap and place in 55-60C waterbath until needed.
- When Agarose has cooled to enough to touch comfortably, add 10microliters Syber Safe. Swirl.
- 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
- Add 2 g per liter coomassie blue for stain. Destain is exactly the same, except do not add coomassie blue.
- 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.
- 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.
