Amersham Biosciences Hoefer DALT, User Manual, Page: 47 / 70

Hoefer DALT System Recipes
Amersham Biosciences
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SDS Electrophoresis Buffer
(25 mM Tris, 192 mM glycine, 0.1% SDS, approximate pH 8.3, 20 liters)
SDS Equilibration Buffer
(50 mM Tris-HCl pH 8.8, 6 M urea, 30% glycerol, 2% SDS, bromophenol blue,
200 ml)
Agarose Sealing Solution
(25 mM Tris, 192 mM glycine, 0.1% SDS, 0.5% agarose, approximate pH 8.3, 25 ml)
Final Conc. Amount
Tris (MW 121.14) 25 mM 60.5 g
Glycine (MW 75.07) 192 mM 288.0 g
SDS (FW 288.38) 0.1% (w/v) 20.0 g
Double distilled H
2
O to 20 liters
Do not adjust the pH of this solution.
Final Conc. Amount
Tris-Cl, pH 8.8 (1.5 M solution) 50 mM 6.67 ml
Urea (FW 60.06) 6 M 72.07 g
Glycerol (87% v/v, MW 92.09) 30% (v/v) 69 ml
SDS (FW 288.38) 2% (w/v) 4.0 g
Bromophenol blue trace a few grains
Double distilled H
2
O to 200 ml
Store at -20 °C.
This is a stock solution. Add DTT or iodoacetamide before using.
Final Conc. Amount
SDS Electrophoresis Buffer (see above) 25 ml
Agarose (NA or M) 125 mg
Bromophenol blue trace a few grains
Combine all ingredients in a 250 ml Erlenmeyer flask. Swirl to disperse. On a low setting, heat
in a microwave oven until the agarose is completely dissolved, about 1 minute. Do not allow the
solution to boil over.
Allow the agarose to cool slightly before using. Do not adjust pH.
The volume of sealing solution prepared from this recipe is sufficient for a complete set of
second-dimension DALT gels.