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Nucleic Acids Res. 1999 Sep 15;27(18):e19.

Polymorphism analysis and gene detection by minisequencing on an array of gel-immobilized primers.

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  • 1Joint Human Genome Program, Engelhardt Institute of Molecular Biology, 32 Vavilov Str., Moscow 117984, Russia.


Two procedures, multibase and multiprimer, have been developed for single nucleotide extension of primers immobilized within polyacrylamide gel pads on a microchip. In the multibase assay, a primer is next to a polymorphic nucleotide; the nucleotide is identified by the specificity with which the primer incorporates fluorescently labeled dideoxyribo-nucleoside triphosphates. In the multiprimer assay, several primers containing different 3'-terminal nucleotides overlapping the variable nucleotide in DNA are used. The polymorphic nucleotide is identified according to the primer that is extended. The methods were compared for diagnosis of beta-thalassemia mutations. Isothermal amplification of the fluorescent signal was achieved by performing both assays at elevated temperature. Anthrax toxin genes were identified in a model system using this amplification method.

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