Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1978 Mar;5(3):653–665. doi: 10.1093/nar/5.3.653

Separation of very large DNA molecules by gel electrophoresis.

W L Fangman
PMCID: PMC342013  PMID: 417296

Abstract

Very large DNA molecules were separated by electrophoresis in horizontal slab gels of dilute agarose. Conditions of electrophoresis were developed using intact DNA molecules from the bacterial viruses lambda, T4 and G. Their DNAs have molecular weights (M) of 32 million, 120 million, and 500 million, respectively. Several electrophoresis conditions were found which give sufficiently high mobilities and large differences that these DNAs are separated in a short time. Electrophoresis in 0.1% agarose at 2.5 V/cm of gel length separates T4 and lambda DNAs by 2.0 cm, and G and T4 DNAs by 1.0 cm in only 10 hr. With some conditions DNA mobilities are directly proportional to log M for M values from 10 to 500 million. The procedures used will allow rapid molecular weight determination and separation of very large DNA molecules.

Full text

PDF
653

Images in this article

664Image
on p.664

Selected References

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

  1. Byers B., Goetsch L. Electron microscopic observations on the meiotic karyotype of diploid and tetraploid Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1975 Dec;72(12):5056–5060. doi: 10.1073/pnas.72.12.5056. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Donelli G., Dore E., Frontali C., Grandolfo M. E. Structure and physico-chemical properties of bacteriophage G. III. A homogeneous DNA of molecular weight 5 times 10(8). J Mol Biol. 1975 Jun 5;94(4):555–565. doi: 10.1016/0022-2836(75)90321-6. [DOI] [PubMed] [Google Scholar]
  3. Dore E., Frontali C., Grignoli M. The molecular complexity of G DNA. Virology. 1977 Jun 15;79(2):442–445. doi: 10.1016/0042-6822(77)90370-1. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. 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]
  6. Henckes G., Crochet M., Labedan B., Legault-Demare J. Fractionation of high molecular-weight duplex DNA molecules on dilute agarose gels. Anal Biochem. 1974 Jul;60(1):1–14. doi: 10.1016/0003-2697(74)90126-2. [DOI] [PubMed] [Google Scholar]
  7. Johnson P. H., Grossman L. I. Electrophoresis of DNA in agarose gels. Optimizing separations of conformational isomers of double- and single-stranded DNAs. Biochemistry. 1977 Sep 20;16(19):4217–4225. doi: 10.1021/bi00638a014. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Mosig G., Carnighan J. R., Bibring J. B., Cole R., Bock H. G., Bock S. Coordinate variation in lengths of deoxyribonucleic acid molecules and head lengths in morphological variants of bacteriophage T4. J Virol. 1972 May;9(5):857–871. doi: 10.1128/jvi.9.5.857-871.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Petes T. D., Byers B., Fangman W. L. Size and structure of yeast chromosomal DNA. Proc Natl Acad Sci U S A. 1973 Nov;70(11):3072–3076. doi: 10.1073/pnas.70.11.3072. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Petes T. D., Fangman W. L. Sedimentation properties of yeast chromosomal DNA. Proc Natl Acad Sci U S A. 1972 May;69(5):1188–1191. doi: 10.1073/pnas.69.5.1188. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  13. Zimm B. H. Anomalies in sedimentation. IV. Decrease in sedimentation coefficients of chains at high fields. Biophys Chem. 1974 Apr;1(4):279–291. doi: 10.1016/0301-4622(74)80014-1. [DOI] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES