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Simultaneous Ferguson plot analysis, using electrophoresis on a single agarose pore gradient gel, of DNA fragments contained in a mixture

A recent study [1] has demonstrated the feasibility of obtaining Ferguson plots in agarose gel electrophoresis, using a single pore gradient gel. We now report three remedies for defects in the previous experimental approach: (i) UV‐absorbing media for density stabilization of the gel is avoided by replacing 5‐(N‐2, 3‐dihydroxypropylacetamido)‐2, 4, 6‐triiodo‐N,N′bis (2, 3‐dihydroxypropyl) isophthalamide (Nycodenz) with heavy water; this renders the method applicable to ethidium bromide‐labeled DNA. (ii) The density stabilizing medium is kept from having an effect on field strength. (iii) Data collection by uninterrupted time‐lapse photography is possible by using an apparatus with a quartz window. These three measures make the method practical for the gel electrophoretic identification and physical characterization of DNA species, potentially up to 50 kb in size.