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. 1976 Aug;73(8):2776–2780. doi: 10.1073/pnas.73.8.2776

Determination of molecular weights by fluctuation spectroscopy: application to DNA.

M Weissman, H Schindler, G Feher
PMCID: PMC430740  PMID: 822421

Abstract

A method for determining molecular weights of macromolecules by measuring spontaneous concentration fluctuations is described. The method is absolute, rapid, and requires no shearing forces on the molecules. We have applied this technique to the determination of molecular weight of DNA molecules. The molecular weight values obtained for T2 phage DNA (1.14 X 10(8)) and replicating Escherichia coli DNA (3.9 X 10(9)) agree with previous results. By monitoring individual molecules, an estimate of the molecular weight of nuclei and individual chromosomal DNA molecules of Drosophila melanogaster was obtained.

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Selected References

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

  1. Bauer W., Vinograd J. The interaction of closed circular DNA with intercalative dyes. I. The superhelix density of SV40 DNA in the presence and absence of dye. J Mol Biol. 1968 Apr 14;33(1):141–171. doi: 10.1016/0022-2836(68)90286-6. [DOI] [PubMed] [Google Scholar]
  2. Canellakis E. S., Shaw Y. H., Hanners W. E., Schwartz R. A. Diacridines: bifunctional intercalators. I. Chemistry, physical chemistry and growth inhibitory properties. Biochim Biophys Acta. 1976 Feb 5;418(3):277–289. doi: 10.1016/0005-2787(76)90290-2. [DOI] [PubMed] [Google Scholar]
  3. Cooper S., Helmstetter C. E. Chromosome replication and the division cycle of Escherichia coli B/r. J Mol Biol. 1968 Feb 14;31(3):519–540. doi: 10.1016/0022-2836(68)90425-7. [DOI] [PubMed] [Google Scholar]
  4. Crothers D. M., Zimm B. H. Viscosity and sedimentation of the DNA from bacteriophages T2 and T7 and the relation to molecular weight. J Mol Biol. 1965 Jul;12(3):525–536. doi: 10.1016/s0022-2836(65)80310-2. [DOI] [PubMed] [Google Scholar]
  5. Feher G., Weissman M. Fluctuation spectroscopy: determination of chemical reaction kinetics from the frequency spectrum of fluctuations. Proc Natl Acad Sci U S A. 1973 Mar;70(3):870–875. doi: 10.1073/pnas.70.3.870. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Freifelder D. Molecular weights of coliphages and coliphage DNA. IV. Molecular weights of DNA from bacteriophages T4, T5 and T7 and the general problem of determination of M. J Mol Biol. 1970 Dec 28;54(3):567–577. doi: 10.1016/0022-2836(70)90127-0. [DOI] [PubMed] [Google Scholar]
  7. Kavenoff R., Zimm B. H. Chromosome-sized DNA molecules from Drosophila. Chromosoma. 1973;41(1):1–27. doi: 10.1007/BF00284071. [DOI] [PubMed] [Google Scholar]
  8. Klotz L. C., Zimm B. H. Size of DNA determined by viscoelastic measurements: results on bacteriophages, Bacillus subtilis and Escherichia coli. J Mol Biol. 1972 Dec 30;72(3):779–800. doi: 10.1016/0022-2836(72)90191-x. [DOI] [PubMed] [Google Scholar]
  9. Le Pecq J. B., Le Bret M., Barbet J., Roques B. DNA polyintercalating drugs: DNA binding of diacridine derivatives. Proc Natl Acad Sci U S A. 1975 Aug;72(8):2915–2919. doi: 10.1073/pnas.72.8.2915. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. Magde D., Elson E. L., Webb W. W. Fluorescence correlation spectroscopy. II. An experimental realization. Biopolymers. 1974 Jan;13(1):29–61. doi: 10.1002/bip.1974.360130103. [DOI] [PubMed] [Google Scholar]
  12. Massie H. R., Zimm B. H. THE USE OF HOT PHENOL IN PREPARING DNA. Proc Natl Acad Sci U S A. 1965 Dec;54(6):1641–1643. doi: 10.1073/pnas.54.6.1641. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. RUBENSTEIN I., THOMAS C. A., Jr, HERSHEY A. D. The molecular weights of T2 bacteriophage DNA and its first and second breakage products. Proc Natl Acad Sci U S A. 1961 Aug;47:1113–1122. doi: 10.1073/pnas.47.8.1113. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Roberts T. M., Lauer G. D., Klotz L. C. Physical studies on DNA from "primitive" eucaryotes. CRC Crit Rev Biochem. 1975;3(4):349–449. [PubMed] [Google Scholar]
  15. Ryder O. A., Smith D. W. Isolation of membrane-associated folded chromosomes from Escherichia coli: effect of protein synthesis inhibition. J Bacteriol. 1974 Dec;120(3):1356–1363. doi: 10.1128/jb.120.3.1356-1363.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Steinkamp J. A., Crissman H. A. Automated analysis of deoxyribonucleic acid, protein and nuclear to cytoplasmic relationships in tumor cells and gynecologic specimens. J Histochem Cytochem. 1974 Jul;22(7):616–621. doi: 10.1177/22.7.616. [DOI] [PubMed] [Google Scholar]
  17. Sueoka N., Yoshikawa H. The chromosome of Bacillus subtilis. I. Theory of marker frequency analysis. Genetics. 1965 Oct;52(4):747–757. doi: 10.1093/genetics/52.4.747. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Waring M. J. Complex formation between ethidium bromide and nucleic acids. J Mol Biol. 1965 Aug;13(1):269–282. doi: 10.1016/s0022-2836(65)80096-1. [DOI] [PubMed] [Google Scholar]

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