, Volume 4, Issue 6, pp 325-330

Reproducibility and correlation study of three deoxyribonucleic acid hybridization procedures

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Reproducibility of relative binding ratios (RBR) was studied with 26 labeled DNA-unlabeled DNA hybridization reactions carried out at two temperatures (60 and 75°C) and following three procedures: the S1-nuclease method with precipitation of S1-resistant DNA by trichloroacetic acid (S1-TCA), the S1-nuclease method with adsorption of S1-resistant DNA onto DE81 filters (S1-DE81), and the hydroxyapatite (HA) method. Analyses of variance indicated that different hybridization methods give different RBR, but similar percent divirgence (ΔT m) results. A curvilinear relationship was found between RBR data obtained with S1-TCA and HA procedures at 60 or 75°C, and between 60°C RBR data obtained with S1-DE81 and HA procedures. At 75°C, RBR obtained with S1-DE81 and HA methods are comparable without any transformation of the data.