Instruction manual

50x TAE Buffer (1 L)
Tris base 242.0 g
Acetic acid, glacial 57.1 ml
0.5 M EDTA, pH 8.0 100.0 ml
Mix and add dH
2
O to 1 L. Autoclave for 20–30 minutes. Store at room temperature.
0% Denaturing Solution (100 ml)
7.5% Gel 10% Gel 12.0% Gel
40% Acrylamide/Bis (37.5:1) 18.8 ml 25.0 ml 30.0 ml
50x TAE buffer 2.0 ml 2.0 ml 2.0 ml
dH
2
O to 100 ml to 100 ml to 100 ml
Degas for about 10–15 minutes. Store at 4 °C in a brown bottle for about 1 month.
100% Denaturing Solution (100 ml)
7.5% Gel 10% Gel 12.0% Gel
40% Acrylamide/Bis (37.5:1) 18.8 ml 25.0 ml 30.0 ml
50x TAE buffer 2.0 ml 2.0 ml 2.0 ml
Formamide (deionized) 40.0 ml 40.0 ml 40.0 ml
Urea 42.0 g 42.0 g 42.0 g
dH
2
O to 100 ml to 100 ml to 100 ml
Degas for about 10–15 minutes. Store at 4 °C in a brown bottle for about 1 month.
For other denaturing solutions, use the volumes in the 100% Denaturing Solution, with the
exception of the formamide and urea. For these reagents, use the volumes indicated below.
Denaturing Solution
10% 20% 30% 40% 50% 60% 70% 80% 90%
Formamide (ml) 4 8 12 16 20 24 28 32 36
Urea (g) 4.2 8.4 12.6 16.8 21 25.2 29.4 33.6 37.8
10% Ammonium Persulfate (1 ml)
Ammonium persulfate 0.1 g
dH
2
O 1.0 ml
Mix and store at -20 °C for about a month.
D GENE Dye Solution (10 ml)
Bromophenol blue 0.05 g
Xylene cyanol 0.05 g
1x TAE buffer 10.0 ml
Store at room temperature.
1x TAE Running Buffer (7 L)
50x TAE buffer 140.0 ml
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