Skip to main content
Biochemical Journal logoLink to Biochemical Journal
. 1969 Jun;113(1):131–138. doi: 10.1042/bj1130131

The determination of the molecular weight of ribonucleic acid by polyacrylamide-gel electrophoresis. The effects of changes in conformation

U E Loening 1
PMCID: PMC1184612  PMID: 4896715

Abstract

1. The effects of changes in experimental conditions on the mobility of RNA in polyacrylamide-gel electrophoresis were investigated. 2. The linear relation between log(molecular weight) and electrophoretic mobility was shown to be independent within limits of salt or gel concentration. 3. The relative mobility of RNA with low content of guanylic acid and cytidylic acid residues was decreased in low-ionic-strength buffer. This was related to a small relative decrease in sedimentation coefficient. 4. However, Mg2+ ion caused almost no increase in mobility although it was associated with large increases in sedimentation coefficient. This suggested opposing actions of Mg2+ ion on the size and effective charge of the RNA. 5. It is concluded that the method provides a satisfactory measurement of molecular weight, which is almost independent of the nucleotide composition of RNA at moderate salt concentrations.

Full text

PDF
131

Selected References

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

  1. Bishop D. H., Claybrook J. R., Spiegelman S. Electrophoretic separation of viral nucleic acids on polyacrylamide gels. J Mol Biol. 1967 Jun 28;26(3):373–387. doi: 10.1016/0022-2836(67)90310-5. [DOI] [PubMed] [Google Scholar]
  2. Bockstahler L. E., Kaesberg P. Isolation and properties of RNA from bromegrass mosaic virus. J Mol Biol. 1965 Aug;13(1):127–137. doi: 10.1016/s0022-2836(65)80084-5. [DOI] [PubMed] [Google Scholar]
  3. Click R. E., Tint B. L. Comparative sedimentation rates of plant, bacterial and animal ribosomal RNA. J Mol Biol. 1967 Apr 14;25(1):111–122. doi: 10.1016/0022-2836(67)90282-3. [DOI] [PubMed] [Google Scholar]
  4. Dingman C. W., Peacock A. C. Analytical studies on nuclear ribonucleic acid using polyacrylamide gel electrophoresis. Biochemistry. 1968 Feb;7(2):659–668. doi: 10.1021/bi00842a022. [DOI] [PubMed] [Google Scholar]
  5. Loening U. E., Ingle J. Diversity of RNA components in green plant tissues. Nature. 1967 Jul 22;215(5099):363–367. doi: 10.1038/215363a0. [DOI] [PubMed] [Google Scholar]
  6. Loening U. E. Molecular weights of ribosomal RNA in relation to evolution. J Mol Biol. 1968 Dec;38(3):355–365. doi: 10.1016/0022-2836(68)90391-4. [DOI] [PubMed] [Google Scholar]
  7. Loening U. E. The fractionation of high-molecular-weight ribonucleic acid by polyacrylamide-gel electrophoresis. Biochem J. 1967 Jan;102(1):251–257. doi: 10.1042/bj1020251. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Parish J. H., Kirby K. S. Reagents which reduce interactions between ribosomal RNA and rapidly labelled RNA from rat liver. Biochim Biophys Acta. 1966 Dec 21;129(3):554–562. doi: 10.1016/0005-2787(66)90070-0. [DOI] [PubMed] [Google Scholar]
  9. Pene J. J., Knight E., Jr, Darnell J. E., Jr Characterization of a new low molecular weight RNA in HeLa cell ribosomes. J Mol Biol. 1968 May 14;33(3):609–623. doi: 10.1016/0022-2836(68)90309-4. [DOI] [PubMed] [Google Scholar]
  10. Penman S. RNA metabolism in the HeLa cell nucleus. J Mol Biol. 1966 May;17(1):117–130. doi: 10.1016/s0022-2836(66)80098-0. [DOI] [PubMed] [Google Scholar]
  11. Petermann M. L., Pavlovec A. The subunits and structural ribonucleic acids of Jensen sarcoma ribosomes. Biochim Biophys Acta. 1966 Feb 21;114(2):264–276. doi: 10.1016/0005-2787(66)90308-x. [DOI] [PubMed] [Google Scholar]
  12. Richards E. G., Coll J. A., Gratzer W. B. Disc electrophoresis of ribonucleic acid in polyacrylamide gels. Anal Biochem. 1965 Sep;12(3):452–471. doi: 10.1016/0003-2697(65)90212-5. [DOI] [PubMed] [Google Scholar]
  13. Stanley W. M., Jr, Bock R. M. Isolation and physical properties of the ribosomal ribonucleic acid of Escherichia coli. Biochemistry. 1965 Jul;4(7):1302–1311. doi: 10.1021/bi00883a014. [DOI] [PubMed] [Google Scholar]
  14. Willems M., Wagner E., Laing R., Penman S. Base composition of ribosomal RNA precursors in the HeLa cell nucleolus: further evidence of non-conservative processing. J Mol Biol. 1968 Mar 14;32(2):211–220. doi: 10.1016/0022-2836(68)90005-3. [DOI] [PubMed] [Google Scholar]

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

RESOURCES