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© IRL Press Limited

Analysis of microdensitometric data in terms of probability of cleavage of DNA

Andrzej Galat * and Irving H. Goldberg

Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School 25 Shattuck Street, Boston, MA 02115, USA

*To whom reprint requests should be sent

Computer-aided analysis of autoradiographic films of DNA fragments is presented. The Powell least-squares procedure is used for optimization of parameters for components of complex densitometric curves. Since each densitometric spectrum may be divided for several non-overlapped blocks of bands, there is no upper limit on the number of parameters which must be optimized. Eight shapes for the component bands are utilized: symmetric and asymmetric Gauss and Cauchy functions, direct, symmetric and asymmetric product of Gauss function and inverse of Cauchy function, and log-normal function. The probability of DNA cleavage is calculated with correction for multiple cuts. The method presented was applied to detailed analysis of densitometric spectra of a 21-bp DNA restriction fragment and allowed for direct correlation between structural micro-heterogeneity of DNA and the resulting cutting pattern. This method should facilitate the analysis of densitometric data from antibiotic-induced cleavage of DNA and footprinting experiments.

Received on May 7, 1987; accepted on July 21, 1987

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