Volume Required For (Ml); Final %T 10% 12.5; Acrylamide Stock 1.5 M Tris-Cl, Ph; Water 10% Sds 10% Aps 10% Temed - GE HEALTHCARE Ettan DALTsix User Manual

Electrophoresis system

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Table of Contents

8 Recipes

Gel storage solution

(0.375 M Tris-Cl, pH 8.8, 0.1% (w/v) SDS, 2.0 l)

Tris-Cl (1.5 M, pH 8.8)

10% (w/v) SDS

Distilled or deionized water

Store at 4 ºC.

10× SDS electrophoresis buffer

(250 mM Tris, 1.92 M glycine, 1.0% (w/v) SDS, approximate pH 8.3, 20 l)

Tris (MW 121.14)

Glycine (MW 75.07)

SDS (MW 288.38)

Distilled or deionized water

Do not adjust the pH of this solution.

SDS equilibration buffer

(50 mM Tris-Cl, pH 8.8, 6 M urea, 30% (v/v) glycerol, 2% (w/v) SDS,

bromophenol blue, 200 ml)

Tris-Cl (1.5 M, pH 8.8)

Urea (MW 60.06)

Glycerol (87% [v/v], MW 92.09)

SDS (MW 288.38)

Bromophenol blue

Distilled or deionized water

Store at -20 ºC. This is a stock solution. Add DTT or iodoacetamide before using.

Agarose sealing solution

(25 mM Tris, 192 mM glycine, 0.1% (w/v) SDS, bromophenol blue,

0.5% (w/v) agarose, 25 ml)

1× SDS electrophoresis buffer (see above)

Agarose (NA or M)

Bromophenol blue

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 com-

pletely melted, about 1 min. Do not allow the solution to boil over. Allow the agarose to cool slightly before using. Do not adjust pH.

Homogeneous gel solutions

600 ml

final %T

Acrylamide stock

1.5 M Tris-Cl, pH 8.8

Water

10% SDS

10% APS

10% TEMED

final conc.

0.375 M

0.1 % (w/v)

final conc.

250 mM

1.92 M

1.0% (w/v)

final conc.

50 mM

6 M

30% (v/v)

2% (w/v)

trace

final conc.

trace