DNA typing with digoxigenin-11-dideoxyuridinetriphosphate-labelled oligonucleotide probes enables non-radioactive analysis using a dual detection system: application for screening HLA-DQA1 polymorphisms.
We describe a standardized, highly sensitive, nonradioactive detection procedure for HLA class-II typing of DQA1 alleles and suballeles which has important and cost-effective application in studying HLA disease associations. The procedure involves polymerase chain reaction-based target DNA sequence amplification and dot blotting followed by stringent hybridization with digoxigenin-11-dideoxyuridine triphosphate 3'-end-labelled allele-specific oligonucleotide probes. The dual detection system described here makes this procedure very versatile.
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Berufsgenossenschaftliches Forschungsinstitut für Arbeitsmedizin, Abteilung Molekulare Genetik, Bochum, FRG.
Thiele
A
A
Perichon
B
B
Krishnamoorthy
R
R
Baur
X
X
D000483:Alleles; D000595:Amino Acid Sequence; D001483:Base Sequence; D003856:Deoxyuracil Nucleotides; D054306:Dideoxynucleotides; D004076:Digoxigenin / Q000031:analogs & derivatives*; D006683:HLA-DQ Antigens / Q000235:genetics*; D059848:HLA-DQ alpha-Chains; D006650:Histocompatibility Testing / Q000379:methods; D006801:Humans; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D011110:Polymorphism, Genetic; D017422:Sequence Analysis, DNA / Q000379:methods
D003856:Deoxyuracil Nucleotides; D054306:Dideoxynucleotides; D006683:HLA-DQ Antigens; D059848:HLA-DQ alpha-Chains; C069050:HLA-DQA1 antigen; D015345:Oligonucleotide Probes; C070581:digoxigenin-11-2',3'-dideoxyuridine 5'-triphosphate; D004076:Digoxigenin