Simultaneous measurements of mobility, dispersion, and orientation of DNA during steady-field gel electrophoresis coupling a fluorescence recovery after photobleaching apparatus with a fluorescence detected linear dichroism setup.
Phys Rev E Stat Phys Plasmas Fluids Relat Interdiscip Topics
; 61(6 Pt B): 6993-8, 2000 Jun.
Article
en En
| MEDLINE
| ID: mdl-11088393
Orientation of molecules is responsible for the loss of separability during steady-field gel electrophoresis. In this work we develop a technique to measure simultaneously the relevant parameters involved in the separation mechanism: electrophoretic mobility, band broadening, and molecular orientation. To do that we have associated a fluorescence recovery after photobleaching (FRAP) apparatus with a fluorescence detected linear dichroism setup. This coupling allows one to follow the buildup of orientation during the FRAP experiment. Because orientation involves a change in the angular distribution of fluorescence, we have added a fluorescence polarization setup which can be used in parallel with the FRAP and gives an exact value of the steady-state orientation factor. We illustrate the possibilities of these combined experiments by analyzing the coupling of electrophoretic transport and orientation of lambda DNA in 1% agarose gels.
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Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
ADN Viral
/
Electroforesis en Gel de Agar
/
Polarización de Fluorescencia
Idioma:
En
Revista:
Phys Rev E Stat Phys Plasmas Fluids Relat Interdiscip Topics
Asunto de la revista:
MEDICINA
Año:
2000
Tipo del documento:
Article
País de afiliación:
Francia
Pais de publicación:
Estados Unidos