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. 1975 May;2(5):647–653. doi: 10.1093/nar/2.5.647

The kinetics of in situ hybridization.

P Szabo, R Elder, Uhlenbeck
PMCID: PMC343617  PMID: 1170554

Abstract

The kinetics of the cytological or in situ DNA-RNA hybridization reaction between 125-I HeLa cell 18S and 28S rRNA and the interphase nuclei of chinese hamster cells were studied. The reaction is consistent with the expected first order kinetics, and the value of the rate constant was very similar to the value obtained from analogous filter disc DNA-RNA hybridization experiments. This similarity in the rate constants and the known relationship between the rate constant and the complexity of the RNA hybridized, define conditions to optimize the in situ hybridization reaction.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Biedler J. L., Riehm H. Cellular resistance to actinomycin D in Chinese hamster cells in vitro: cross-resistance, radioautographic, and cytogenetic studies. Cancer Res. 1970 Apr;30(4):1174–1184. [PubMed] [Google Scholar]
  2. Birnstiel M. L., Sells B. H., Purdom I. F. Kinetic complexity of RNA molecules. J Mol Biol. 1972 Jan 14;63(1):21–39. doi: 10.1016/0022-2836(72)90519-0. [DOI] [PubMed] [Google Scholar]
  3. Flavell R. A., Birfelder E. J., Sanders J. P., Borst P. DNA-DNA hybridization on nitrocellulose filters. 1. General considerations and non-ideal kinetics. Eur J Biochem. 1974 Sep 16;47(3):535–543. doi: 10.1111/j.1432-1033.1974.tb03722.x. [DOI] [PubMed] [Google Scholar]
  4. Franklin R. M. Purification and properties of the replicative intermediate of the RNA bacteriophage R17. Proc Natl Acad Sci U S A. 1966 Jun;55(6):1504–1511. doi: 10.1073/pnas.55.6.1504. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gall J. G., Pardue M. L. Formation and detection of RNA-DNA hybrid molecules in cytological preparations. Proc Natl Acad Sci U S A. 1969 Jun;63(2):378–383. doi: 10.1073/pnas.63.2.378. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hutton J. R., Wetmur J. G. Effect of chemical modification on the rate of renaturation of deoxyribonucleic acid. Deaminated and glyoxalated deoxyribonucleic acid. Biochemistry. 1973 Jan 30;12(3):558–563. doi: 10.1021/bi00727a032. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Prensky W., Steffensen D. M., Hughes W. L. The use of iodinated RNA for gene localization. Proc Natl Acad Sci U S A. 1973 Jun;70(6):1860–1864. doi: 10.1073/pnas.70.6.1860. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Steffensen D. M., Wimber D. E. Hybridization of nucleic acids to chromosomes. Results Probl Cell Differ. 1972;3:47–63. doi: 10.1007/978-3-540-37149-6_5. [DOI] [PubMed] [Google Scholar]
  10. Wimber D. E., Steffensen D. M. Localization of gene function. Annu Rev Genet. 1973;7:205–223. doi: 10.1146/annurev.ge.07.120173.001225. [DOI] [PubMed] [Google Scholar]

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