Partial denaturation mapping of phage T4 DNA at low temperature

Abstract

The partial-denaturation map of T4 DNA is obtained by using benzyldodecyldimethyl ammonium chloride in the presence of a high concentration of formamide. In this way suitable conditions for preparation of electron microscope specimens and partial denaturation within a temperature range low enough to minimize the endonucleolytic cleavage, are realized. It is found that, under our experimental conditions, the denaturation increase depends mainly on the appearance of new denaturation sites rather than on the lengthening of DNA segments already denatured. Because of the DNA circular permutation it is necessary to align the measured maps to obtain the denaturation pattern. This is done through a computer program and informations on the distribution of the regions with highest (A-T) content along the genome are obtained. The results of contour length measurements of lambda and T4 DNA's are also reported.