Coomassie (quick stain)

This protocol was obtained from Megan in the Doudna lab. Quick, no-destain-required Coomassie protocol:

Recipe:

0.25 g Coomassie Brilliant Blue G250

900mL H2O

100 mL EtOH

2.5 mL conc HCl or 5 mL 6M HCl


 * Stir overnight @ room temp, store @ RT.

Before staining, gels have to be boiled twice in water to get rid of SDS. Heat the gel in 5-10 mL of stain in the microwave for 30 seconds, then shake at room temperature. Bands should appear in <5 min on a clear background, wait 15 min for nice bands. No destaining necessary.

So in a nutshell:

Boil 2x in H2O

Microwave in stain/30 sec

Shake @ RT/15 min or longer

Pour off stain, add H2O/pour off until stain is gone

Look at your beautiful bands!

An alternative quick stain protocol from Jorge:
Stain = 2.5g comomassie + 500mL EtOH + 400mL H2O + 100mL Acetic Acid De-stain = Same as stain sans dye

Here is an alternate method for staining and destaining, involving the use of the microwave:

1. Place gel in coomassie stain (enough to cover it) and microwave it until it boils (~1min) 2. Allow it to cool down (~20min) 3. Replace the staining solution with the de-staining solution, and microwave until it boils (~1min)4. Let it cool down