A simple logarithmic amplifier for the photographic quantitation of DNA in gel electrophoresis |
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Authors: | W Hörz KV Oefele H Schwab |
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Institution: | Institut für Physiologische Chemie, Physikalische Biochemie und Zellbiologie der Universität, Goethestrasse 33, 8000 Munich 2, Federal Republic of Germany |
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Abstract: | For the quantitative determination of nonradioactive DNA fragments by gel electrophoresis, it is usually necessary to photograph the gel after staining with ethidium bromide and evaluate the negative by densitometry. It has previously been shown that, because of the logarithmic nature of the photographic process, it is not the optical density (E) of the film which is proportional to the amount of DNA in the gel but instead the value , γ being a film constant. We describe the design of a simple instrument that converts E into . The instrument can be built in any electronic workshop at low cost. When it is used together with a standard recording densitometer, densitometric tracings of are obtained directly. These tracings can be quantitated by simple peak area measurements, thereby circumventing complicated mathematical transformations. Quantitative analyses of a linear and an exponential densitogram of restriction nuclease digested plasmid DNA are presented to demonstrate the usefulness of the instrument. |
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