## Abstract Polyacrylamide gel electrophoresis (PAGE) is used frequently for isolation and purification of DNA fragments. In the present study, DNA fragments extracted from polyacrylamide gels showed significant band broadening in capillary electrophoresis (CE). A pHY300PLK (a shuttle vector functi
Diffusion, Joule heating, and band broadening in capillary gel electrophoresis of DNA
β Scribed by Dr. Gary W. Slater; Pascal Mayer; Paul D. Grossman
- Publisher
- John Wiley and Sons
- Year
- 1995
- Tongue
- English
- Weight
- 872 KB
- Volume
- 16
- Category
- Article
- ISSN
- 0173-0835
No coin nor oath required. For personal study only.
β¦ Synopsis
Diffusion, Joule heating, and band broadening in capillary gel electrophoresis of DNA
We calculated the longitudinal and transverse diffusion coefficients for a DNA molecule undergoing gel electrophoresis in the limit where it is reptating through a dense polymer matrix. Our results indicate that both diffusion coefficients increase with the electric field intensity. The transverse and longitudinal diffusion coefficients are roughly equal for regular field intensities, but the former dominates for the high field intensities normally used for capillary gel electrophoresis (CE). This has important implications for the optimization of CE. Our results clearly show that the naive use of the zero-field diffusion constant, the Einstein relation or the longitudinal diffusion constant when calculating the contribution of the parabolic temperature profile to band broadening may lead to large overestimates under typical CE conditions. Finally, we show that the field-dependent diffusion coefficients may be responsible for the existence of an optimal field intensity for CE, even if Joule heating is neglected.
π SIMILAR VOLUMES
## Abstract Diffusion coefficients of oligonucleotides have been measured in polyacrylamide gel capillaries without application of an electrical field. A relationship between such diffusion coefficients and molecular size of the oligonucleotides could be achieved. The slower diffusion of larger oli
## Abstract Using a computer simulation algorithm based on the reptation model, we investigated the effects of pulsed field gel electrophoresis on the separation of singleβstranded DNA molecules in denaturing polyacrylamide gels. Pulsed fields that combine two different field intensities were found