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Detection of beta-thalassemia mutations by ASO hybridization of PCR amplified DNA with digoxigenin ddUTP labeled oligonucleotides.

Abstract
A simple procedure for nonradioactive labeling of oligonucleotides has recently been developed (1). It consists of 3' end labeling of oligonucleotides with terminal transferase by incorporation of a single digoxigenin labeled dideoxy uridine triphosphate. We used these oligonucleotides for allele specific oligomer hybridization of polymerase chain reaction amplified DNA, followed by an enzyme-linked immunoassay and subsequent enzyme-catalyzed color reaction. We compared this procedure with the standard radioactive oligonucleotide hybridization technique through the detection of the most common Mediterranean beta-thalassemia mutations. This procedure was also used for the confirmation of a new mutation at position -87 (C----A) (2) of the beta-globin gene and for the subsequent family analysis.
AuthorsD G Efremov, A J Dimovski, G D Efremov
JournalHemoglobin (Hemoglobin) Vol. 15 Issue 6 Pg. 525-33 ( 1991) ISSN: 0363-0269 [Print] England
PMID1814858 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
Chemical References
  • Dideoxynucleotides
  • Oligodeoxyribonucleotides
  • Uracil Nucleotides
  • 2',3'-dideoxyuridine-5'-triphosphate
  • Globins
  • DNA
  • DNA Nucleotidylexotransferase
  • Digoxigenin
Topics
  • Alleles
  • Colorimetry
  • DNA (genetics)
  • DNA Mutational Analysis
  • DNA Nucleotidylexotransferase
  • Dideoxynucleotides
  • Digoxigenin
  • Enzyme-Linked Immunosorbent Assay
  • Globins (genetics)
  • Humans
  • Nucleic Acid Hybridization
  • Oligodeoxyribonucleotides
  • Polymerase Chain Reaction
  • Thalassemia (genetics)
  • Uracil Nucleotides

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