Molecular Hybridization and Its Application to RNA Tumor Virus Research
The remarkable fact that, under appropriate conditions, the two strands of native DNA can be dissociated and reassociated in vitro (Doty et al., 1960; Marmur and Lane, 1960) supplied a use-full tool for the examination of genetic homologies. The formation of hybrid molecules could be demonstrated from the DNA of two different bacterial species (Schildkraut et al., 1961a) or viruses (Schildkraut et al., 1962). This basic methodology was soon extended to the formation of RNA-DNA hybrid molecules (Hall and Spiegelman, 1960; Schildkraut et al., 1961b). In order to prevent the competition between the hybridization process and the reannealing of DNA in the reaction medium, techniques were developed for the immobilization of single-stranded DNA in cellulose (Bautz and Hall, 1962), in agar (Bolton and Mc Carthy, 1962) and on nitrocellulose filters (Nygaard and Hall, 1964; Gillespie and Spiegelman, 1965). So it became possible to measure the extent of reassociation of radioactivity labeled single-stranded DNA or RNA with the immobilized DNA.
KeywordsMolecular Hybridization Reassociation Kinetic Reiteration Frequency Viral Genome Equivalent Endogenous Reverse Transcriptase
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