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 |
| 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.
Agarose Gels
Recommended Agarose Percentage for Size Separation
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."
| % 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."