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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1969 Jun;63(2):378–383. doi: 10.1073/pnas.63.2.378

FORMATION AND DETECTION OF RNA-DNA HYBRID MOLECULES IN CYTOLOGICAL PREPARATIONS*

Joseph G Gall 1, Mary Lou Pardue 1
PMCID: PMC223575  PMID: 4895535

Abstract

A technique is described for forming molecular hybrids between RNA in solution and the DNA of intact cytological preparations. Cells in a conventional tissue squash are immobilized under a thin layer of agar. Next they are treated with alkali to denature the DNA and then incubated with tritium-labeled RNA. The hybrids are detected by autoradiography. The technique is illustrated by the hybridization of ribosomal RNA to the amplified ribosomal genes in oocytes of the toad Xenopus. A low level of gene amplification was also detected in premeiotic nuclei (oogonia).

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Selected References

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  1. BAUTZ E. K., HALL B. D. The isolation of T4-specific RNA on a DNA-cellulose column. Proc Natl Acad Sci U S A. 1962 Mar 15;48:400–408. doi: 10.1073/pnas.48.3.400. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. BOLTON E. T., McCARTHY B. J. A general method for the isolation of RNA complementary to DNA. Proc Natl Acad Sci U S A. 1962 Aug;48:1390–1397. doi: 10.1073/pnas.48.8.1390. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brown D. D., Dawid I. B. Specific gene amplification in oocytes. Oocyte nuclei contain extrachromosomal replicas of the genes for ribosomal RNA. Science. 1968 Apr 19;160(3825):272–280. doi: 10.1126/science.160.3825.272. [DOI] [PubMed] [Google Scholar]
  4. CONGER A. D., FAIRCHILD L. M. A quick-freeze method for making smear slides permanent. Stain Technol. 1953 Nov;28(6):281–283. doi: 10.3109/10520295309105555. [DOI] [PubMed] [Google Scholar]
  5. Evans D., Birnstiel M. L. Localization of amplified ribosomal DNA in the oocyte of Xenopus laevis. Biochim Biophys Acta. 1968 Aug 23;166(1):274–276. doi: 10.1016/0005-2787(68)90517-0. [DOI] [PubMed] [Google Scholar]
  6. Flamm W. G., McCallum M., Walker P. M. The isolation of complementary strands from a mouse DNA fraction. Proc Natl Acad Sci U S A. 1967 Jun;57(6):1729–1734. doi: 10.1073/pnas.57.6.1729. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gall J. G. Differential synthesis of the genes for ribosomal RNA during amphibian oögenesis. Proc Natl Acad Sci U S A. 1968 Jun;60(2):553–560. doi: 10.1073/pnas.60.2.553. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gillespie D., Spiegelman S. A quantitative assay for DNA-RNA hybrids with DNA immobilized on a membrane. J Mol Biol. 1965 Jul;12(3):829–842. doi: 10.1016/s0022-2836(65)80331-x. [DOI] [PubMed] [Google Scholar]
  9. HALL B. D., SPIEGELMAN S. Sequence complementarity of T2-DNA and T2-specific RNA. Proc Natl Acad Sci U S A. 1961 Feb 15;47:137–163. doi: 10.1073/pnas.47.2.137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. KIT S. Equilibrium sedimentation in density gradients of DNA preparations from animal tissues. J Mol Biol. 1961 Dec;3:711–716. doi: 10.1016/s0022-2836(61)80075-2. [DOI] [PubMed] [Google Scholar]
  11. NYGAARD A. P., HALL B. D. FORMATION AND PROPERTIES OF RNA-DNA COMPLEXES. J Mol Biol. 1964 Jul;9:125–142. doi: 10.1016/s0022-2836(64)80095-4. [DOI] [PubMed] [Google Scholar]
  12. Sambrook J., Westphal H., Srinivasan P. R., Dulbecco R. The integrated state of viral DNA in SV40-transformed cells. Proc Natl Acad Sci U S A. 1968 Aug;60(4):1288–1295. doi: 10.1073/pnas.60.4.1288. [DOI] [PMC free article] [PubMed] [Google Scholar]

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