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      Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia.

      Science (New York, N.Y.)

      Polymorphism, Genetic, Anemia, Sickle Cell, diagnosis, genetics, Base Sequence, Clinical Laboratory Techniques, DNA Restriction Enzymes, DNA-Directed DNA Polymerase, Alleles, diagnostic use, Escherichia coli, Gene Amplification, Globins, Humans, Nucleic Acid Hybridization

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          Abstract

          Two new methods were used to establish a rapid and highly sensitive prenatal diagnostic test for sickle cell anemia. The first involves the primer-mediated enzymatic amplification of specific beta-globin target sequences in genomic DNA, resulting in the exponential increase (220,000 times) of target DNA copies. In the second technique, the presence of the beta A and beta S alleles is determined by restriction endonuclease digestion of an end-labeled oligonucleotide probe hybridized in solution to the amplified beta-globin sequences. The beta-globin genotype can be determined in less than 1 day on samples containing significantly less than 1 microgram of genomic DNA.

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          2999980

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