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DYE MIGRATION: POLYACRYLAMIDE DENATURING GELS |
Dyes will migrate to the same point as double-stranded DNA of the
indicated size in a denaturing polyacrylamide gel.
| Gel % |
Bromophenol Blue |
Xylene Cyanol |
| 5.0 |
35bp |
140bp |
| 6.0 |
26bp |
106bp |
| 8.0 |
19bp |
75bp |
| 10.0 |
12bp |
55bp |
| 20.0 |
8bp |
28bp |
Adapted from Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989) In:
Molecular Cloning: A Laboratory Manual, Cold Spring Harbor
Laboratory, Cold Spring Harbor, NY. |
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DYE MIGRATION: POLYACRYLAMIDE NONDENATURING GELS |
Dyes will migrate to the same point as double-stranded DNA of the
indicated size in a nondenaturing polyacrylamide gel.
| Gel % |
Bromophenol Blue |
Xylene Cyanol |
| 3.5 |
100bp |
460bp |
| 5.0 |
65bp |
260bp |
| 8.0 |
45bp |
160bp |
| 12.0 |
20bp |
70bp |
| 15.0 |
15bp |
60bp |
| 20.0 |
12bp |
45bp |
Adapted from Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989) In:
Molecular Cloning: A Laboratory Manual, Cold Spring Harbor
Laboratory, Cold Spring Harbor, NY. |
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DYE MIGRATION IN 0.5-1.4% AGAROSE GELS |
Dyes will migrate to the same point as double-stranded DNA of the
indicated size in an agarose gel. Sizes are approximate.
| GoTaq® blue dye |
4kb |
| Xylene cyanol FF |
4kb |
| Bromophenol Blue |
300bp |
| Orange G |
50bp |
| GoTaq® yellow dye |
10bp |
Some information adapted from Sambrook, J., Fritsch, E.F. and
Maniatis, T. (1989) In: Molecular Cloning: A Laboratory Manual,
Cold Spring Harbor Laboratory, Cold Spring Harbor, NY. |
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GEL PERCENTAGES: RESOLUTION OF LINEAR DNA ON AGAROSE
GELS |
Recommended
% Agarose |
Optimum Resolution
for Linear DNA
(Size of fragments in
base pairs) |
| 0.5 |
1,000–30,000 |
| 0.7 |
800–12,000 |
| 1.0 |
500–10,000 |
| 1.2 |
400–7,000 |
| 1.5 |
200–3,000 |
| 2.0 |
50–2,000 |
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GEL PERCENTAGES: RESOLUTION OF PROTEINS ON
POLYACRYLAMIDE GELS |
Recommended
% Acrylamide |
Protein Size Range |
| 8 |
40–200kDa |
| 10 |
21–100kDa |
| 12 |
10–40kDa |
Dalton (Da) is an alternate name for the atomic mass unit and kilodalton
(kDa) is 1,000 daltons. Thus a peptide with a mass of 64kDa has a
molecular weight of 64,000 grams per mole. |
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