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5.9 Stock
solutions
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B. Sample buffer
0.050 mol/l Tris-HAc pH 7.5
Dissolve 0.3 g Tris in 40 ml distilled water. Carefully adjust to pH 7.5 with
HAc (approximately 0.14 ml). Make up to 50 ml with distilled water. Add
0.4 g SDS and a few grains of Bromophenol Blue. Immediately before use
add 40 mg of DTT.
C. Fixing solution
Ethanol
Acetic acid, HAc
Make up to 1000 ml with distilled water.
D. Incubation solution
Ethanol
Sodium acetate
Glutaraldehyde (25% w/v)
Sodium thiosulphate, Na
Make up to 250 ml with distilled water.
E. Silver solution
Silver nitrate
Formaldehyde
Make up to 250 ml with distilled water.
F. Developing solution
Sodium carbonate
Formaldehyde
Make up to 250 ml with distilled water.
G. Stop solution
EDTA-Na
x 2H
O
2
2
Make up to 250 ml with distilled water.
H. Preserving solution
Glycerol (87% w/w)
Make up to 250 ml with distilled water.
I. Destaining solution
Ethanol
Acetic acid
Make up to 1000 ml with distilled water.
K. Coomassie solution
PhastGel Blue R
Make up to 400 ml with destaining solution.
Heat to 60 °C, stirring constantly, and filter before use.
L. Preserving solution
Glycerol (87% w/w)
Make up to 250 ml with destaining solution.
N. Fixing solution
Trichloroacetic acid
Make up to 500 ml with distilled water.
400 ml
100 ml
75 ml
17.00 g
1.25 ml
S
O
x 5H
O
0.50 g
2
2
3
2
0.25 g
50 µl
6.25 g
25 µl
3.65 g
25 ml
250 ml
80 ml
1 tablet
25 ml
100 g
5. Operation
47
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