Bio-Rad DCode™ Universal Mutation Detection System - Manual (Page 50)

40% Acrylamide/Bis Solutions (1x TBE)
Reagent
6% Gel 8% Gel 10% Gel 12% Gel
40% Acrylamide/Bis 6 ml 8 ml 10 ml 12 ml
10x TBE (see note) 4 ml 4 ml 4 ml 4 ml
Urea (optional) see note see note see note see note
TEMED 40 µl 40 µl 40 µl 40 µl
10% Ammonium persulfate 400 µl 400 µl 400 µl 400 µl
Total volume 40 ml 40 ml 40 ml 40 ml
Add dH
2
O to 40 ml and mix. Cast the gel immediately after adding the TEMED and ammonium
persulfate.
Note: For 0.5x TBE, add 2 ml.
For 15% urea, add 6 gm.
2x DEM Solution (0.6x TBE)
Reagent
1x Gel 0.8x Gel
2x DEM 20 ml 16 ml
10x TBE (see note) 2.4 ml 2.4 ml
Urea (optional) see note see note
TEMED 40 µl 40 µl
10% Ammonium persulfate 400 µl 400 µl
Total volume 40 ml 40 ml
Add dH
2
O to 40 ml and mix. Cast the gel immediately after adding the TEMED and ammonium
persulfate.
Note:
For 0.5x TBE, add 2 ml.
For 1x TBE, add 4 ml.
For 15% urea, add 6 gm.
10% Ammonium Persulfate
Reagent
Amount
Ammonium persulfate 0.1 g
dH
2
O 1.0 ml
Store at –20°C for about a 1 week.
2x Gel Loading Dye
Reagent
Amount Final Concentration
2% Bromophenol blue 0.25 ml 0.05%
2% Xylene cyanol 0.25 ml 0.05%
100% Glycerol 7.0 ml 70%
dH
2
O 2.5 ml
Total volume 10.0 ml
Store at room temperature.
1x TBE Running Buffer
Reagent
2 L Buffer 7 L Buffer
10x TBE buffer 200 ml 700 ml
dH
2
O 1,800 ml 6,300 ml
0.6x TBE Running Buffer
Reagent
2 L Buffer 7 L Buffer
10x TBE buffer 120 ml 420 ml
dH
2
O 1,880 ml 6,580 ml
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