NONENZYMATIC LABELING OF GENE PROBES FOR DIAGNOSIS OF SICKLE-CELL MUTATION AND BETA-THALASSEMIA

作者：BURGI W^*; KELLER E; SCHMIDHEINI T

来源：Schweizerische Medizinische Wochenschrift, 1992, 122(42): 1567-1570.

A method is described in which allele-specific oligonucleotides (ASO) are labeled non-enzymatically with digoxigenin (DIG). The ASOs were synthesized in a DNA synthesizer. A primary C6-alkylamine was incorporated at the 5'end of the ASO. The derivative was coupled to digoxigenin-3-O-methylcarbonyl-epsilon-aminocaproic acid-N-hydroxy-succinimide ester. The DIG-ASO probes were isolated from the crude preparation and hybridized with amplified genomic DNA. The annealed probes were detected with an ELISA test. The DIG-ASO probes were successfully used for the identification of sickle cell disease genotypes and the most common beta-thalassemia mutations.

• 出版日期1992-10-17