Enzymatic Amplification of β-Globin Genomic Sequences and Restriction Site Analysis for Diagnosis of Sickle Cell Anemia
Randall K. Saiki, Stephen Scharf, Fred Faloona, Kary B. Mullis, Glenn T. Horn, Henry A. Erlich, Norman Arnheim
Summary
This paper reported the first published application of in vitro primer-mediated enzymatic amplification of DNA—the technique that became known as the polymerase chain reaction (PCR)—as part of a rapid, sensitive prenatal diagnostic test for sickle cell anemia. The authors amplified specific β-globin target sequences from genomic DNA roughly 220,000-fold and then distinguished the normal (βA) and sickle (βS) alleles by restriction endonuclease digestion of a hybridized end-labeled oligonucleotide probe. The combined procedure allowed genotyping in under a day using far less than one microgram of genomic DNA.
Key findings
- Demonstrated primer-mediated enzymatic amplification of specific β-globin genomic sequences, achieving an ~220,000-fold increase in target DNA copies—the first published use of what became PCR.
- Developed an oligonucleotide restriction assay (solution hybridization plus restriction digestion) to discriminate the βA and βS alleles for diagnosis of sickle cell anemia.
- Showed the combined method could determine genotype in less than one day using substantially less than 1 µg of genomic DNA, enabling rapid molecular diagnosis.
Subjects & keywords
Cite this paper
Randall K. Saiki, Stephen Scharf, Fred Faloona, Kary B. Mullis, Glenn T. Horn, Henry A. Erlich, & Norman Arnheim (1985). Enzymatic Amplification of β-Globin Genomic Sequences and Restriction Site Analysis for Diagnosis of Sickle Cell Anemia. Science. https://doi.org/10.1126/science.2999980
@article{saiki1985enzymatic,
author = {Randall K. Saiki and Stephen Scharf and Fred Faloona and Kary B. Mullis and Glenn T. Horn and Henry A. Erlich and Norman Arnheim},
title = {Enzymatic Amplification of β-Globin Genomic Sequences and Restriction Site Analysis for Diagnosis of Sickle Cell Anemia},
journal = {Science},
year = {1985},
doi = {10.1126/science.2999980},
url = {https://doi.org/10.1126/science.2999980}
}