|
Polyacrylamide Gels
Recommended Polyacrylamide
% for Separation in Denaturing Gels
|
% Acrylamide
|
Effective Range of
Separation (nucleotides)
|
|
3.5
|
>500
|
|
5
|
151-500
|
|
10
|
61-150
|
|
15
|
30-60
|
|
20
|
<30
|
Dye Migration in Denaturing
Polyacrylamide Gels
|
% Acrylamide
|
Xylene Cyanol (nucleotides)
|
Bromophenol Blue (nucleotides)
|
|
5
|
130
|
35
|
|
6
|
106
|
26
|
|
8
|
75
|
19
|
|
10
|
55
|
12
|
|
12
|
28
|
8
|
Adapted from Sambrook, J., Fritsch, E.F., and
Maniatis, T. (1989) in: Molecular Cloning: A Laboratory Manual,
Cold Springs Harbor Laboratory.
Dye Migration in Nondenaturing
Polyacrylamide Gels
|
% Acrylamide
|
Effective Range of
Separation (bp)
|
Xylene Cyanol (nucleotides)
|
Bromophenol Blue (nucleotides)
|
|
3.5
|
100-1000
|
460
|
100
|
|
5
|
100-500
|
260
|
65
|
|
8
|
60-400
|
160
|
45
|
|
12
|
50-200
|
70
|
20
|
|
15
|
25-150
|
60
|
15
|
|
20
|
5-100
|
45
|
12
|
Adapted from Sambrook, J., Fritsch, E.F., and
Maniatis, T. (1989) in: Molecular Cloning: A Laboratory Manual,
Cold Springs Harbor Laboratory.
Agarose Gels
Recommended Agarose Percentage for Size Separation
|
% Agarose
|
Size Fragments Separated
(bp)
|
|
0.5
|
1,000 to 30,000
|
|
0.7
|
800 to 12,000
|
|
1.0
|
500 to 10,000
|
|
1.2
|
400 to 7,000
|
|
1.5
|
200 to 3,000
|
|
3-4 (sieving agarose)
|
10 to 1000
|
Adapted from Ausubel, F.M., Brent, R., Kingston,
R.E., Moore, D.D., Seidman, J.G., Smith, J.A., Struhl, K. (1995)
in "Current Protocols in Molecular Biology."
|