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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 |
| 20 | 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.
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 1,000 |
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."
仅供科研使用，不可用于诊断目的。