Observation of DNA molecules undergoing capillary electrophoresis.
This paper presents a method to observe the motions and configurations of large DNA molecules undergoing capillary electrophoresis (CE). A simple device to perform CE horizontally under microscopic observation is designed and images of single DNA molecules inside the capillary are obtained using an epi-fluorescence microscope. DNA molecules moved towards the negative electrode when an electric field was applied. The mobilities of three types of DNA (T4 and lambda bacteriophage DNA and PBR322 plasmid DNA) were measured at different electric field strength. The mobility vs. electric field strength curves of these three large DNAs showed that the mobility remained constant at high electric field strength (200-600 Volt/cm) and increased significantly at low electric field strength (less than or equal to 50 Volt/cm.). The apparent mobilities of the large DNA molecules were independent of molecular weight. At electric field strengths greater than or equal to 400 Volt/cm., big aggregates (snowballs) of DNA molecules formed and moved upstream towards the positive electrode. When the field was turned off, the aggregates dissociated into a cloud of single DNA molecules, and diffused into the solution.
Song
L
L
Cell and Molecular Biology Division, Lawrence Berkeley Laboratory, Berkeley, CA 94720.
Maestre
M F
MF
D004247:DNA / Q000737:chemistry*; D004279:DNA, Viral / Q000737:chemistry; D004563:Electrochemistry; D004586:Electrophoresis / Q000379:methods*; D007091:Image Processing, Computer-Assisted; D008856:Microscopy, Fluorescence; D009690:Nucleic Acid Conformation; D013604:T-Phages / Q000235:genetics; D013816:Thermodynamics
D004279:DNA, Viral; D004247:DNA