Urea gel - preparation of urea gel (Dec/13/2005 )

Hi all
i am working with very low molecular weight proteins so i am trying to work on urea gel, but the problem is that when i use 8M urea in gel, i get very bad gel. I have tried the gel without urea thats some what better.
Is anybody knows the exact composition of urea gel, and the role of high concentration of urea in gel.

thanx

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OK, here goes!

For a 10% gel, use 10 mL of acrylamide/bis solution, 3.5 mL of 10XTBE (tris-borate-edta standard buffer), urea is 15 grams and 10 mL of water. Microwave until clear, avoid boiling. Degas for about 2 minutes which will also allow to cool slightly. Add 0.3 mL of 10% ammonium persulfate and 35 uL of TEMED (I add at same time). Swirl and pour.

Good Luck!

Cindy

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i'd better go for warming in 60-70°C waterbath, because sometimes in mircowave the solution might start boiling, which leads to very nasty contamination of the mircowave by acrylamid. that should be avoided for sure ... besides that ... agreed

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