5% Acrylamide; 7.5M Urea
Note: Urea and acrylamide are toxic, so wear gloves and avoid skin contamination
1X TBE
250ml
40% acrylamide
62.5 mL (Biorad 40% acrylamide/bis solution 19:1 catalog#1610144)
Urea
227.29 g (Sigma U-1250/5kg)
Mix acrylamide and TBE
Add urea and stir until it dissolves (at least 3 hours).
Note
: Do
not
heat the solution to speed up the dissolution of urea since this will produce uric acid due to the breakdown of urea.
Bring to 500ml volume in graduated cylinder
Filter with 0.45um filter into 1L glass bottle
Transfer solution from glass bottles to brown plastic bottles
Store at 4°C
in the dark
.