[Non-enzymatic marking of gene probes using digoxigenin for the diagnosis of sickle cell mutation and beta thalassemia].
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-3-0-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.
Bürgi
W
W
Zentrallaboratorium, Kantonsspital Aarau.
Keller
E
E
Schmidheini
T
T
D000328:Adult; D000755:Anemia, Sickle Cell / Q000235:genetics*; D001483:Base Sequence; D002648:Child; D002675:Child, Preschool; D004076:Digoxigenin; D005260:Female; D006455:Hemoglobins, Abnormal / Q000235:genetics; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D015345:Oligonucleotide Probes; D017086:beta-Thalassemia / Q000235:genetics*
D006455:Hemoglobins, Abnormal; D015345:Oligonucleotide Probes; D004076:Digoxigenin
Nicht-enzymatische Markierung von Gen-Sonden mit Digoxigenin für die Diagnose der Sichelzellmutation und der beta-Thalassämie.