Detection of Specific DNA Sequences — The Southern Blot

  • Christopher G. P. Mathew


In a previous chapter (Chapter 14) the characterisation of DNA by restriction analysis on agarose gels has been described. In such cases the DNA sample would normally be about one microgram of a single type of DNA sequence. However, if you wished to analyse the structure of a specific gene from a complex organism without prior purification of the gene, you would have to detect picogram amounts of a single type of DNA sequence among about a million other types. The method used would therefore have to be very sensitive and very specific. This exacting task can be accomplished by means of a technique devised by Edwin Southern of Edinburgh University.1 The technique has come to be known as the Southern transfer or Southern blot, and has had an enormous impact on molecular biology and genetics.


Southern Blot Sickle Cell Anaemia Globin Gene Nitrocellulose Filter Antenatal Diagnosis 
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Further Reading

  1. 1.
    P. Chambon, ‘Split genes’, Sci. Amer., 244 (1981), 48–59CrossRefGoogle Scholar
  2. 2.
    B.D. Hall. L. Haar and K. Klepp, ‘Development of the nitrocellulose filter technique for RNA-DNA hybridisation’, Trends in Biochemical Sciences, 5 (1980), 254–256CrossRefGoogle Scholar
  3. 3.
    P.F.R. Little, ‘Ante-natel diagnosis of haemoglobinopathies’ in R. Williamson (ed.), Genetic Engineering, (Academic Press. 1981 ) vol. 1, 61-102.Google Scholar


  1. 1.
    E.M. Southern. ‘Detection of Specific Sequences among DNA fragments separated by gel electrophoresis’. J. Mol. Biol., 98 (1975). 503–17.PubMedCrossRefGoogle Scholar
  2. 2.
    J.C. Alwine, D.J. Kemp, B.A. Parker, J. Reiser, J. Renart, G.R. Stark and G.M. Wahl, ‘Detection of specific RNAs or specific fragments of DNA by fractionation in gels and transfer to diazobenzyloxymethyl paper’ in R. Wu. (ed.). Methods in Enzymology, 68 (1979), 220–42.PubMedCrossRefGoogle Scholar
  3. 3.
    Southern, ‘Detection of specific sequences among DNA fragments’.Google Scholar
  4. 4.
    E.M. Southern. ‘Gel electrophoresis of restriction fragments’ in R. Wu, (ed.). Methods in Enzymology, 68 (1979), 152–76.PubMedCrossRefGoogle Scholar
  5. 5.
    G.M. Wahl, M. Stern and G.R. Stark, ‘Efficient transfer of large DNA fragments from agarose gels to DBM paper and rapid hybridisation using dextran sulphate’, Proc. Natl. Aca. Sci. USA, 76 (1979), 3683–7.CrossRefGoogle Scholar
  6. 6.
    M. Bittner, P. Kupferer and C.F. Morris. ‘Electrophoretic transfer of proteins and nucleic acids from slab gels to diazobenzyloxymethyl cellulose or nitrocellulose sheets’, Anal. Biochem., 102 (1980), 459–71.PubMedCrossRefGoogle Scholar
  7. 7.
    O. Faub, S. Bratosin, M. Horowitz and Y. Alom, ‘The initiation of transcription of SV40 DNA at late time after infection’, Virology, 92 (1979), 310–23.CrossRefGoogle Scholar
  8. 8.
  9. 9.
    Bittner, Kupferer and Morris, ‘Electrophoretic transfer’.Google Scholar
  10. 10.
    Alwine, Kemp, Parker, Reiser, Renart, Stark and Wahl, ‘Detection of specific RNAs’.Google Scholar
  11. 11.
    D.T. Denhardt, ‘A membrane-filter technique for the detection of complementary DNA’, Biochem. Biophys. Res. Comm., 23 (1966), 641–6.PubMedCrossRefGoogle Scholar
  12. 12.
    M. Barinaga, R. Franco, J. Meinkoth, E. Ong and G.M. Wahl, (1981) ‘Methods for transfer of DNA, RNA and protein to nitrocellulose paper and diazotized paper solid supports’. Schleicher and Schuell technical publication No. 352-4.Google Scholar
  13. 13.
    Wahl, Stern and Stark, ‘Efficient transfer of large DNA fragments’.Google Scholar
  14. 14.
    R. A. Laskey, ‘The use of intensifying screens or organic scintillators for visualizing radioactive molecules resolved by gel electrophoresis’ in L. Grossman and K. Moldave (eds.). Methods in Enzymology, 65 (1980), part 1, 363–71.PubMedCrossRefGoogle Scholar
  15. 15.
    R.A. Laskey and A.D. Mills, ‘Enhanced autoradiographic detection of 32P and 125I using intensifying screens and hypersensitized film’, FEBS Letts, 82 (1977), 314–8.CrossRefGoogle Scholar
  16. 16.
  17. 17.
    Southern, ‘Gel electrophoresis’.Google Scholar
  18. 18.
    D. Gillespie, ‘The formation and detection of DNA-RNA hybrids’ in L. Grossman and K. Moldave (eds.), Methods in Enzymology, 12B (1968), 641–68.CrossRefGoogle Scholar
  19. 19.
    Barinaga, Franco, Meinkoth, Ong and Wahl, ‘Methods for transfer’.Google Scholar
  20. 20.
    Laskey, ‘The use of intensifying screens’.Google Scholar
  21. 21.
  22. 22.
    Alwine, Kemp, Parker, Reiser, Renart, Stark and Wahl, ‘Detection of specific RNAs’.Google Scholar
  23. 23.
  24. 24.
    Barinaga, Franco, Meinkoth, Ong and Wahl, ‘Methods for transfer’.Google Scholar
  25. 25.
    R. Breathnach, J.L. Mandel, P. Chambon, ‘Ovalbumin gene is split in chicken DNA’, Nature, 270 (1977), 314–19.PubMedCrossRefGoogle Scholar
  26. 26.
    M. Ehrlich and R. Wang, ‘5-Methylcytosine in eukaryotic DNA’, Science, 212 (1981), 1350–7.PubMedCrossRefGoogle Scholar
  27. 27.
    J. Stalder, M. Groudine, J.B. Dodgson, J.D. Engel and H. Weintraub, ‘Hb switching in chicken’, Cell, 19 (1980), 973–80.PubMedCrossRefGoogle Scholar
  28. 28.
    Southern, ‘Detection of specific sequences among DNA fragments’.Google Scholar
  29. 29.
    T. Maniatis, H. Fritsch, J. Lauer and R. Lawn, ‘The molecular genetics of human haemoglobins’, Ann. Rev. Genet., 14 (1980), 145–78.PubMedCrossRefGoogle Scholar
  30. 30.
    R.T. Geever, L.B. Wilson, F.S. Nallaseth, P.P. Milner, M. Bittner and J.T. Wilson, ‘Direct identification of sickle cell anaemia by blot hybridisation’, Proc. Natl. Acad. Sci. USA, 78 (1981), 5081–5.PubMedCrossRefGoogle Scholar
  31. 31.
    Y.W. Kan and A.M. Dozy, ‘Ante-natal diagnosis of sickle cell anaemia by DNA analysis of amniotic fluid cells’, The Lancet, ii (1978), 910–2.Google Scholar
  32. 32.
    D. Botstein, R.L. White, M. Skolnick and R. Davis, ‘Construction of a genetic linkage map in man using restriction fragment length polymorphisms’, Amer. J. Hum. Genet., 32 (1980), 314–31.PubMedGoogle Scholar
  33. 33.
    M. Denaro, H. Blanc, M. Johnson, K. Chen, E. Wilmsen, L. Cavalli-Sforza and D. Wallace, ‘Ethnic variation in Hpa 1 endonuclease cleavage patterns of human mitochondrial DNA’, Proc. Natl. Acad. Sci. USA., 78 (1981) 5768–2.PubMedCrossRefGoogle Scholar

Copyright information

© John M. Walker and Wim Gaastra 1983

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  • Christopher G. P. Mathew

There are no affiliations available

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