Ligation assembly and detection of polynucleotides on...

Details Ligation assembly and detection of polynucleotides on... Ligation assembly and detection of polynucleotides on...

C - Chemistry, Metallurgy

12

Q

C12Q 1/68 (2006.01) C12P 19/34 (2006.01)

Patent

CA 2353076

The present invention concerns methods of assembly of a polynucleotide on a solid-support by performing steps of annealing, ligation, and extension. The steps may be repeated in a cyclical manner to assemble immobilized double- or single-stranded polynucleotides with functional gene properties. The immobilized polynucleotides may be amplified by the polymerase chain reaction, and detected and quantitated by an exonuclease assay with a self-quenching, fluorescent probe. The polynucleotide may be cleaved from the solid-support by chemical or enzymatic cleavage.

L'invention concerne des procédés de liaison d'un polynucléotide sur un support solide, à savoir par l'annellation, la ligature et l'extension. Les étapes peuvent être répétées de manière cyclique jusqu'à lier les polynucléotides immobilisés à simple ou double brin présentant des propriétés génétiques fonctionnelles. Les polynucléotides immobilisés peuvent être amplifiés par la réaction en chaîne de la polymérase, décelés et quantifiés par un test d'exonucléase présentant une sonde fluorescente à désactivation automatique. Le polynucléotide peut être coupé du support solide par clivage chimique ou enzymatique.

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