Purification of Plant Ribosomal DNA
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The general features of isopycnic centrifugation of DNA and the separation of main-band from satellite DNAs in solutions of cesium chloride are discussed in the preceeding chapter. This chapter describes a modification of the procedure that enhances the separation of specific DNA sequences from main-band DNA. The method is based on the fact that actinomycin D binds preferentially to DNA molecules with a high-GC content. This lowers their density relative to main-band and other satellite DNAs so that they can be resolved separately. Cesium chloride-actinomycin D gradients have been used to isolate sea urchin histone genes (1) and plant ribosomal DNA (2), and to separate genes coding for plant 25S and 18S from 5S RNA genes (3) and as a prelude to the cloning of ribosomal DNA (4).
KeywordsSatellite DNAs Nitrocellulose Filter Gradient Fraction Cesium Chloride Isopycnic Centrifugation
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