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J Virol. 1974 August; 14(2): 225-230
Copyright © 1974 American Society for Microbiology. All Rights Reserved.

Reliability of the RNA-DNA Filter Hybridization for the Detection of Oncornavirus-Specific DNA Sequences

The University of California School of Medicine, Department of Microbiology and Immunology, Los Angeles, California 90024

ABSTRACT

Denatured DNA from leukemic myeloblasts or uninfected chicken embryos, immobilized on nitrocellulose filters, was hybridized to a vast excess of [³H]70S RNA from purified avian myeloblastosis virus. The viral RNA was eluted from the RNA-DNA hybrids, purified, and then rehybridized in solution to an excess of either leukemic or normal chicken embryonic DNA. This study revealed that all the slow and the fast hybridizing viral RNA sequences detectable by liquid hybridization in DNA excess had hybridized to the filter bound DNA. Both techniques also gave similar values for the number of 28S ribosomal RNA genes contained in a chicken cell genome: 210 by the liquid hybridization procedure and 218 by the filter hybridization technique. Therefore, filter hybridization can accurately detect DNA sequences present in relatively few numbers in the genome of higher organisms.

This article has been cited by other articles:

• McClements, W, Skalka, A. (1977). Analysis of chicken ribosomal RNA genes and construction of lambda hybrids containing gene fragments. Science 196: 195-197 [Abstract]