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.
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Authors | D G Efremov, A J Dimovski, G D Efremov |
Journal | Hemoglobin
(Hemoglobin)
Vol. 15
Issue 6
Pg. 525-33
( 1991)
ISSN: 0363-0269 [Print] England |
PMID | 1814858
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Chemical References |
- Dideoxynucleotides
- Oligodeoxyribonucleotides
- Uracil Nucleotides
- 2',3'-dideoxyuridine-5'-triphosphate
- Globins
- DNA
- DNA Nucleotidylexotransferase
- Digoxigenin
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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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