Abstract
Allelic sequence variation has been analysed by synthetic oligonucleotide hybridization probes which can detect single base substitutions in human genomic DNA1–3. An allele-specific oligonucleotide (ASO) will only anneal to sequences that match it perfectly, a single mismatch being sufficient to prevent hybridization under appropriate conditions4. To improve the sensitivity, specificity and simplicity of this approach, we used the polymerase chain reaction (PCR) procedure5 to enzymatically amplify a specific segment of the β-globin or HLA-DQα gene in human genomic DNA before hybridization with ASOs. This in vitro amplification method, which produces a >105-fold increase in the amount of target sequence, permits the analysis of allelic variation with as little as 1 ng of genomic DNA and the use of a simple ‘dot blot’ for probe hybridization. As a further simplification, PCR amplification has been performed directly on crude cell lysates, eliminating the need for DNA purification.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 51 print issues and online access
$199.00 per year
only $3.90 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
Conner, B. J. et al. Proc. natn. Acad. Sci. U.S.A. 80, 278–272 (1983).
Kidd, V. J., Wallace, R. B., Itakura, K. & Woo, S. L. C. Nature 304, 230–234 (1983).
Pirastu, M. et al. New Engl. J. Med. 309, 284–287 (1983).
Wallace, R. B. et al. Nucleic Acids Res. 6, 3543–3557 (1979).
Faloona, F. A. & Mullis, K. B. Meth. Enzym., (in the press).
Kan, Y. W. & Dozy, A. M. Science 209, 388–391 (1980).
Saiki, R. K. et al. Science 230, 1350–1354 (1985).
Studencki, A. B. et al. Am. J. hum. Genet. 37, 42–51 (1985).
Tuan, D. et al. Proc. natn. Acad. Sci. U.S.A. 80, 6937–6941 (1983).
Scharf, S., Horn, G. & Erlich, H. A. Science 223, 1076–1078 (1986).
Saiki, R. K., Arnheim, N. & Erlich, H. A. Biotechnology 3, 1008–1012 (1985).
Bugawan, T., Horn, G., Long, C. & Erlich, H. A. Manuscript in preparation).
Horn, G. et al. (manuscript in preparation).
Chang, H., Moriuchi, T. & Silver, J. Nature 305, 813–815 (1983).
Moriuchi, J., Moriuchi, T. & Silver, J. Proc. natn. Acad. Sci. U.S.A. 82, 3420–3424 (1985).
Albert, E. D., Baur, M. P. & Mayr, W. R., Histocompatibllity Testing, 4–8 (Springer, Berlin, 1984).
Reed, K. C. & Mann, D. A. Nucleic Acids Res. 13, 7207–7221 (1985).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Saiki, R., Bugawan, T., Horn, G. et al. Analysis of enzymatically amplified β-globin and HLA-DQα DNA with allele-specific oligonucleotide probes. Nature 324, 163–166 (1986). https://doi.org/10.1038/324163a0
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1038/324163a0
This article is cited by
-
Human parvovirus B19 infection in malignant and benign tissue specimens of different head and neck anatomical subsites
Infectious Agents and Cancer (2023)
-
In vitro PCR verification that lysozyme inhibits nucleic acid replication and transcription
Scientific Reports (2023)
-
Prevalence of Human Papillomavirus and Mouse Mammary Tumor Virus Like DNAs in Tumors from Moroccan Breast Cancer Patients
Indian Journal of Gynecologic Oncology (2022)
-
Auto-qPCR; a python-based web app for automated and reproducible analysis of qPCR data
Scientific Reports (2021)
-
The frequency of high-risk human papillomavirus types, HPV16 lineages, and their relationship with p16INK4a and NF-κB expression in head and neck squamous cell carcinomas in Southwestern Iran
Brazilian Journal of Microbiology (2021)
Comments
By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.