Isolation, extraction, and fluorometric measurements of nucleic acids and DNA photorepair in marine macrophytes Public Deposited

http://ir.library.oregonstate.edu/concern/graduate_thesis_or_dissertations/zc77st05q

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  • Nucleic acid estimates in tissues of marine macrophytes with the fluorochromes Hoechst 33258 and ethidium homodimer were severely impaired by high concentrations of interfering compounds. The isolation and extraction techniques used pointed towards compounds of low molecular weight and of polar characteristics that were insensitive to organic solvent extraction and enzymatic degradation. Interference was therefore thought to primarily arise from high concentrations of phenols commonly found in many seaweeds, which might prevent successful fluorochrome binding by complex formation with nucleic acids. None of the investigated isolation methods was sufficiently effective to allow accurate estimates of total nucleic acid content or to reliably establish the metabolic status of seaweeds by RNA/DNA ratios. Despite the uncertainties involved in nucleic acid estimations, DNA isolated by size exclusion chromatography was successfully used to show preliminary evidence for photo-activated DNA repair in marine macrophytes.
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