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. 1977 May;4(5):1409–1418. doi: 10.1093/nar/4.5.1409

The quantitation of fluorescence by photography.

D E Pulleyblank, M Shure, J Vinograd
PMCID: PMC343764  PMID: 302439

Abstract

A method based on theory has been developed for the photographic quantitation of fluorescent substances. DNA stained with ethidium in agarose gels is used as an example of an application of this method. In the course of developing this method we have demonstrated that the empirical methods employed by others authors can give rise to large systematic errors. We have also developed an approximate method based on photographic theory, avoiding the use of digital integration which is required by the rigorous method.

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Selected References

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  1. Brack C., Eberle H., Bickle T. A., Yuan R. A map of the sites on bacteriophage PM2 DNA for the restriction endonucleases HindIII and HpaII. J Mol Biol. 1976 Jun 14;104(1):305–309. doi: 10.1016/0022-2836(76)90016-4. [DOI] [PubMed] [Google Scholar]
  2. DeLeys R. J., Jackson D. A. Electrophoretic analysis of covalently closed SV40 DNA: Boltzmann distributions of DNA species. Nucleic Acids Res. 1976 Mar;3(3):641–652. doi: 10.1093/nar/3.3.641. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Depew D. E., Wang J. C. Conformational fluctuations of DNA helix. Proc Natl Acad Sci U S A. 1975 Nov;72(11):4275–4279. doi: 10.1073/pnas.72.11.4275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dugaiczyk A., Boyer H. W., Goodman H. M. Ligation of EcoRI endonuclease-generated DNA fragments into linear and circular structures. J Mol Biol. 1975 Jul 25;96(1):171–184. doi: 10.1016/0022-2836(75)90189-8. [DOI] [PubMed] [Google Scholar]
  5. Laskey R. A., Mills A. D. Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. Eur J Biochem. 1975 Aug 15;56(2):335–341. doi: 10.1111/j.1432-1033.1975.tb02238.x. [DOI] [PubMed] [Google Scholar]
  6. LePecq J. B., Paoletti C. A fluorescent complex between ethidium bromide and nucleic acids. Physical-chemical characterization. J Mol Biol. 1967 Jul 14;27(1):87–106. doi: 10.1016/0022-2836(67)90353-1. [DOI] [PubMed] [Google Scholar]
  7. Morgan A. R., Paetkau V. A fluorescence assay for DNA with covalently linked complementary sequences. Can J Biochem. 1972 Feb;50(2):210–216. doi: 10.1139/o72-028. [DOI] [PubMed] [Google Scholar]
  8. Polisky B., McCarthy B. Location of histones on simian virus 40 DNA. Proc Natl Acad Sci U S A. 1975 Aug;72(8):2895–2899. doi: 10.1073/pnas.72.8.2895. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Pulleyblank D. E., Shure M., Tang D., Vinograd J., Vosberg H. P. Action of nicking-closing enzyme on supercoiled and nonsupercoiled closed circular DNA: formation of a Boltzmann distribution of topological isomers. Proc Natl Acad Sci U S A. 1975 Nov;72(11):4280–4284. doi: 10.1073/pnas.72.11.4280. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Sharp P. A., Sugden B., Sambrook J. Detection of two restriction endonuclease activities in Haemophilus parainfluenzae using analytical agarose--ethidium bromide electrophoresis. Biochemistry. 1973 Jul 31;12(16):3055–3063. doi: 10.1021/bi00740a018. [DOI] [PubMed] [Google Scholar]
  11. Shure M., Vinograd J. The number of superhelical turns in native virion SV40 DNA and minicol DNA determined by the band counting method. Cell. 1976 Jun;8(2):215–226. doi: 10.1016/0092-8674(76)90005-2. [DOI] [PubMed] [Google Scholar]

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