Abstract
A simple and rapid method for the detection of specific polymorphic restriction sites has been developed that utilizes restriction endonuclease cleavage of a synthetic, end-labeled oligonucleotide probe annealed in solution to the target genomic DNA sequence. The specificity of this method, which depends on the ability of a base pair mismatch within the restriction site to abolish or inhibit cleavage, has been demonstrated by the analysis of genomic DNAs from normal individuals, sickle-cell anemia patients, and sickle-cell carriers (heterozygotes).
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Kan, Y.W. and Dozy, A.M. 1978. Polymorphisms of DNA sequence adjacent to human β-globin structural gene: Relationship to sickle mutation. Proc. Natl. Acad. Sci. USA 75: 5631–5635.
Geever, R.F., Wilson, L.B., Nallaseth, F.S., Milner, P.F., Bittner, M. and Wilson, J.T. 1981. Direct identification of sickle cell anemia by blot hybridization. Proc. Natl. Acad. Sci. USA 78: 5081–5085.
Wilson, J.T., Milner, P.F., Summer, M.E., Nallaseth, F.S., Fadel, H.E., Reindollar, R.H., McDonough, P.G. and Wilson, L.B. 1982. Use of restriction endonucleases for mapping the allele for βS globin. Proc. Natl. Acad. Sci. USA 79: 3628–3631.
Chang, J.C. and Kan, Y.W. 1982. A sensitive new prenatal test for sickle cell anemia. New Engl. J. Med. 707: 30–32.
Orkin, S.H., Little, P.F.R., Kazazian, H.H., Jr. and Boehm, C.D. 1982. Improved detection of the sickle cell mutation by DNA analysis. New Eng. J. Med. 307 32–36.
Davies, K.E. and Ellis, D.P. 1985. An appraisal of the application of recombinant DNA techniques to chromosome defects. Biochem. J. 226: 1–11.
Weatherall, D.J. 1982. The New Genetics and Clinical Practice Neuffield Provincial Hospitals Trust, London.
Conner, B.J., Reyes, A.A., Morin, C., Itakura, K., Teplitz, R.L. and Wallace, B. 1983. Detection of sickle cell βS-globin allele by hybridization with synthetic oligonucleotides. Proc. Natl. Acad. Sci. USA 80: 278–282.
Kidd, V.J., Wallace, R.B., Itakura, K. and Woo, S.L.C. 1983. α1 antitrypsin deficiency detection by direct analysis of the mutation in the gene. Nature 304: 230–234.
Orkin, S.H., Markham, A.F. and Kazazian, H.H., Jr. 1983. Direct detection of the common Mediterranean β-thalassemia gene with synthetic DNA probes: An alternate approach for prenatal diagnosis. J. Clin. Invest. 71: 775–779.
Pirastu, M., Kan, Y.W., Cao, A., Conner, B.J., Teplitz, R.L. and Wallace, R.B. 1983. Prenatal diagnosis of β-thalassemia. New Eng. J. Med. 309: 284–287.
Dove, W.F. and Davidson, N. 1962. Cation effects on the denaturation of DNA. J. Mol. Biol. 5: 467–478.
Tuan, D., Feingold, E., Newman, M., Weissman, S.M. and Forget, B.G. 1983. Different 3′ end points of deletions causing δβ-thalassemia and hereditary persistence of fetal hemoglobin: Implications for the control of γ globin gene expression in man. Proc. Natl. Acad. Sci. USA 80: 6937–6941.
Auffray, C., Lillie, J.W., Arnot, D., Grossberger, D., Kappes, D. and Strominger, J.L. 1984. Isotypic and allotypic variation of human class II histocompatibility antigen α-chain genes. Nature 308: 327–333.
Trowsdale, J., Young, J.A.T., Kelly, A.P., Austin, P.J., Carson, S., Meunier, H., So, A., Erlich, H.A., Spielman, R.S., Bodmer, J. and Bodmer, W.F. 1985. Structure, sequence and polymorphism in the HLA-D region. Immunological Rev., in press.
Collins, F.S., Stoeckert, C.J., Jr., Serjeant, G.R., Forget, B.G. and Weissman, S.M. 1984. Gγβ+ hereditary persistence of fetal hemoglobin: Cosmid cloning and identification of a specific mutation 5′ to the Gγ gene. Proc. Natl. Acad. Sci. USA 81: 4894–4898.
Lawn, R.M., Fritsch, E.F., Parker, R.C., Blake, G. and Maniatis, T. 1978. The isolation and characterization of linked δ- and β-globin genes from a cloned library of human DNA. Cell 15: 1157–1174.
Maniatis, T., Fritsch, E.F. and Sambrook, J. 1982. Molecular Cloning, Cold Spring Harbor Laboratory.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Saiki, R., Arnheim, N. & Erlich, H. A Novel Method for the Detection of Polymorphic Restriction Sites by Cleavage of Oligonucleotide Probes: Application to Sickle-Cell Anemia. Nat Biotechnol 3, 1008–1012 (1985). https://doi.org/10.1038/nbt1185-1008
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1038/nbt1185-1008
