Application of higher derivative techniques to analysis of high-resolution thermal denaturation profiles of reassociated repetitive DNA.

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RESUMO

We have analyzed high-resolution denaturation profiles of reassociated repetitive DNA sequences by using a combination of higher derivative analysis and curve-fitting techniques. Procedures originally used for resolution of components in complex absorption spectra were found to be applicable to high-resolution analysis of melting profiles of reassociated repetitive DNA sequences from pea DNA. Under conditions that eliminate the base composition effect on thermal stability (2.4 M tetraethylammonium chloride), such an anlysis can distinquish "thermal classes" of repetitive DNA duplexes exhibiting different amounts of base pair mismatch. Only a single thermal class is observed in reassociated Escherichia coli DNA whereas at least five classes can be reproducibly distinguished in pea and mung bean DNAs.

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