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A method which facilitates the ordering of DNA restriction fragments

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Summary

The Southern transfer technique has been used to provide a generally applicable method for ordering DNA restriction fragments. It involves electrophoresis of partially digested DNA, transfer to nitrocellulose filter paper and annealing to a 32P-labelled fragment. Only those partials containing that particular fragment will reanneal to the probe and produce bands on autoradiography. The size of each partial in the labelled set is the sum of the size of the fragment used as probe and of one or more adjacent fragments. Thus the size of the adjacent fragments can be determined from the size increments of this set of partials. The method is illustrated by the mapping of certain BamHI sites on coliphage 186 DNA.

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References

  • Denhardt, D.T.: A membrane-filter technique for the detection of complementary DNA. Biochem. Biophys. Res. Commun. 23, 641–646 (1966)

    Google Scholar 

  • Laskey, R.A., Mills, A.D.: Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. Eur. J. Biochem. 56, 335–341 (1975)

    Google Scholar 

  • Rigby, P.W.J., Dieckmann, M., Rhodes, C., Berg, P.: Labelling DNA to high specific activity in vitro by nick translation with DNA polymerase I. J. Mol. Biol. 113, 237–251 (1977)

    Google Scholar 

  • Saint, R.B., Egan, J.B.: A restriction cleavage map of coliphages 186 and P2. Mol. Gen. Genet. 171, 79–89 (1979)

    Google Scholar 

  • Smith, H.O., Birnstiel, M.L.: A simple method for DNA restriction site mapping. Nucleic Acids Res 3, 2387–2398 (1976)

    Google Scholar 

  • Southern, E.M.: Detection of specific sequences among DNA fragments separated by gel electrophoresis. J. Mol. Biol. 98, 503–517 (1975)

    Google Scholar 

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Communicated by W. Arber

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Saint, R.B., Barry Egan, J. A method which facilitates the ordering of DNA restriction fragments. Molec. Gen. Genet. 171, 103–106 (1979). https://doi.org/10.1007/BF00274020

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  • DOI: https://doi.org/10.1007/BF00274020

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