Skip to main content
NCBI home page
Biophysical Journal logoLink to Biophysical Journal
. 1965 Mar;5(2):231–246. doi: 10.1016/s0006-3495(65)86713-3

Chromatographic Separation of Annealed and Enzymatically Synthesized RNA-DNA Hybrids

Marie N Hayashi, M Hayashi, S Spiegelman
PMCID: PMC1367720  PMID: 14268956

Abstract

A procedure is described for the chromatographic detection and isolation of DNA-RNA hybrids on columns of methylated albumin coated on kieselguhr (MAK). Its use is illustrated with both annealed and enzymatically synthesized hybrids. The method has the advantage of a wide range in capacity and resolution and permits actual isolation of the hybrid structure. It is uniquely effective in experiments involving hybridization with small DNA fragments.

Full text

PDF
231

Selected References

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

  1. BASSEL A., HAYASHI M., SPIEGELMAN S. THE ENZYMATIC SYNTHESIS OF A CIRCULAR DNA-RNA HYBRID. Proc Natl Acad Sci U S A. 1964 Sep;52:796–804. doi: 10.1073/pnas.52.3.796. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  3. 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]
  4. CHAMBERLIN M., BERG P. Deoxyribo ucleic acid-directed synthesis of ribonucleic acid by an enzyme from Escherichia coli. Proc Natl Acad Sci U S A. 1962 Jan 15;48:81–94. doi: 10.1073/pnas.48.1.81. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. CHAMBERLIN M., BERG P. MECHANISM OF RNA POLYMERASE ACTION: FORMATION OF DNA-RNA HYBRIDS WITH SINGLE-STRANDED TEMPLATES. J Mol Biol. 1964 Feb;8:297–313. doi: 10.1016/s0022-2836(64)80139-x. [DOI] [PubMed] [Google Scholar]
  6. CHIPCHASE M. I., BIRNSTIEL M. L. ON THE NATURE OF NUCLEOLAR RNA. Proc Natl Acad Sci U S A. 1963 Dec;50:1101–1107. doi: 10.1073/pnas.50.6.1101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. GEIDUSCHEK E. P., NAKAMOTO T., WEISS S. B. The enzymatic synthesis of RNA: complementary interaction with DNA. Proc Natl Acad Sci U S A. 1961 Sep 15;47:1405–1415. doi: 10.1073/pnas.47.9.1405. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. GIACOMONI D., SPIEGELMAN S. Origin and biologic individuality of the genetic dictionary. Science. 1962 Dec 21;138(3547):1328–1331. doi: 10.1126/science.138.3547.1328. [DOI] [PubMed] [Google Scholar]
  9. GOODMAN H. M., RICH A. Formation of a DNA-soluble RNA hybrid and its relation to the origin, evolution, and degeneracy of soluble RNA. Proc Natl Acad Sci U S A. 1962 Dec 15;48:2101–2109. doi: 10.1073/pnas.48.12.2101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. GROS F., GILBERT W., HIATT H. H., ATTARDI G., SPAHR P. F., WATSON J. D. Molecular and biological characterization of messenger RNA. Cold Spring Harb Symp Quant Biol. 1961;26:111–132. doi: 10.1101/sqb.1961.026.01.016. [DOI] [PubMed] [Google Scholar]
  11. GROSSMAN L., LEVINE S. S., ALLISON W. S. The reaction of formaldehyde with nucleotides and T2 bacteriophage DNA. J Mol Biol. 1961 Feb;3:47–60. doi: 10.1016/s0022-2836(61)80007-7. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. HARUNA I., NOZU K., OHTAKA Y., SPIEGELMAN S. AN RNA "REPLICASE" INDUCED BY AND SELECTIVE FOR A VIRAL RNA: ISOLATION AND PROPERTIES. Proc Natl Acad Sci U S A. 1963 Nov;50:905–911. doi: 10.1073/pnas.50.5.905. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. HAYASHI M., HAYASHI M. N., SPIEGELMAN S. DNA CIRCULARITY AND THE MECHANISM OF STRAND SELECTION IN THE GENERATION OF GENETIC MESSAGES. Proc Natl Acad Sci U S A. 1964 Feb;51:351–359. doi: 10.1073/pnas.51.2.351. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. HAYASHI M., HAYASHI M. N., SPIEGELMAN S. RESTRICTION OF IN VIVO GENETIC TRANSCRIPTION TO ONE OF THE COMPLEMENTARY STRANDS OF DNA. Proc Natl Acad Sci U S A. 1963 Oct;50:664–672. doi: 10.1073/pnas.50.4.664. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. HAYASHI M., HAYASHI M. N., SPIEGELMANS Replicating form of a single-stranded DNA virus: isolation and properties. Science. 1963 Jun 21;140(3573):1313–1316. doi: 10.1126/science.140.3573.1313. [DOI] [PubMed] [Google Scholar]
  17. HAYASHI M., SPIEGELMAN S. The selective synthesis of informational RNA in bacteria. Proc Natl Acad Sci U S A. 1961 Oct 15;47:1564–1580. doi: 10.1073/pnas.47.10.1564. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. KANO-SUEOKA T., SPIEGELMAN S. Evidence for a nonrandom reading of the genome. Proc Natl Acad Sci U S A. 1962 Nov 15;48:1942–1949. doi: 10.1073/pnas.48.11.1942. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. MANDELL J. D., HERSHEY A. D. A fractionating column for analysis of nucleic acids. Anal Biochem. 1960 Jun;1:66–77. doi: 10.1016/0003-2697(60)90020-8. [DOI] [PubMed] [Google Scholar]
  20. NYGAARD A. P., HALL B. D. A method for the detection of RNA-DNA complexes. Biochem Biophys Res Commun. 1963 Jul 18;12:98–104. doi: 10.1016/0006-291x(63)90242-0. [DOI] [PubMed] [Google Scholar]
  21. SCHERRER K., LATHAM H., DARNELL J. E. Demonstration of an unstable RNA and of a precursor to ribosomal RNA in HeLa cells. Proc Natl Acad Sci U S A. 1963 Feb 15;49:240–248. doi: 10.1073/pnas.49.2.240. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. SINSHEIMER R. L., LAWRENCE M. IN VITRO SYNTHESIS AND PROPERTIES OF A PHI-X DNA-RNA HYBRID. J Mol Biol. 1964 Feb;8:289–296. doi: 10.1016/s0022-2836(64)80138-8. [DOI] [PubMed] [Google Scholar]
  23. SUEOKA N., CHENG T. Y. Fractionation of nucleic acids with the methylated albumin column. J Mol Biol. 1962 Mar;4:161–172. doi: 10.1016/s0022-2836(62)80048-5. [DOI] [PubMed] [Google Scholar]
  24. YANKOFSKY S. A., SPIEGELMAN S. Distinct cistrons for the two ribosomal RNA components. Proc Natl Acad Sci U S A. 1963 Apr;49:538–544. doi: 10.1073/pnas.49.4.538. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. YANKOFSKY S. A., SPIEGELMAN S. The identification of the ribosomal RNA cistron by sequence complementarity. I. Specificity of complex formation. Proc Natl Acad Sci U S A. 1962 Jun 15;48:1069–1078. doi: 10.1073/pnas.48.6.1069. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. YANKOFSKY S. A., SPIEGELMAN S. The identification of the ribosomal RNA cistron by sequence complementarity. II. Saturation of and competitive interaction at the RNA cistron. Proc Natl Acad Sci U S A. 1962 Aug;48:1466–1472. doi: 10.1073/pnas.48.8.1466. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Biophysical Journal are provided here courtesy of The Biophysical Society

RESOURCES