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Simultaneous Ferguson plot analysis, using electrophoresis on a single agarose pore gradient gel, of DNA fragments contained in a mixture
Author(s) -
Gombocz Erich,
Chrambach Andreas
Publication year - 1991
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150121202
Subject(s) - ethidium bromide , agarose , gel electrophoresis of nucleic acids , electrophoresis , chromatography , agarose gel electrophoresis , pulsed field gel electrophoresis , gel electrophoresis , chemistry , dna , temperature gradient gel electrophoresis , analytical chemistry (journal) , biochemistry , genotype , gene , 16s ribosomal rna
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.