Berglund:Gel Dryer Method
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Low Toxicity Ethanol-based Coomassie Blue staining (from http://micro.nwfsc.noaa.gov/protocols/PAGE-destain.html)
Materials
Stain:
0.25g Coomassie Blue R-250
100ml ethanol
100ml water
stir until dye has dissolved (about 1 hr), add:
25ml glacial acetic acid
25ml water
Destain:
400ml ethanol
100ml glacial acetic acid
500ml water
Methods
- take gel apart in water
- cut off top of gel (stacking part of gel and wells)
- rinse in water a couple of times to get rid of SDS
- shake in 25ml of destain for 5-10 min then pour off destain
- add 50ml of stain (can be reused)
- stain for 30min- 1hr (stain can be heated briefly in microwave to hasten staining process)
- pour off stain
- rinse with water a couple of times
- add destain (25-50ml)
- stop destain with water when bands are quite visible
- put on whatman paper and dry in gel dryer for about 15 min