Transverse agarose pore gradient gel electrophoresis of DNA.
Transverse agarose pore gradient gels were prepared on GelBond in the concentration range of nominally 0.2-1.5% SeaKem GTG agarose, using density stabilization by glycerol and incorporation of a dye to define the gel concentration at each point on the pore gradient gel. The distribution of the dye was evaluated by photography, video-acquisition and digitization of the gradient mixture and by densitometry of the gel. The gel was applied to the electrophoresis of a 1-kb standard ladder of DNA fragments, using standard submarine apparatus. The method extends to agarose gel electrophoresis the benefits of semi-automated analysis of 'Ferguson curves' described in application to polyacrylamide gel by Wheeler et al. (J. Biochem. Biophys. Methods 24, 171-180).
Fawcett
J S
JS
Section on Macromolecular Analysis, National Institute of Child Health and Human Development, NIH, Bethesda, MD 20892.
Wheeler
D
D
Chrambach
A
A
D004396:Coloring Agents; D004247:DNA / Q000032:analysis*; D003720:Densitometry; D004587:Electrophoresis, Agar Gel; D010781:Photography; D014741:Video Recording
D004396:Coloring Agents; D004247:DNA