Enzymatic ligation of 3'-amino-substituted oligonucleotides

Details Enzymatic ligation of 3'-amino-substituted oligonucleotides Enzymatic ligation of 3'-amino-substituted oligonucleotides

C - Chemistry, Metallurgy

12

Q

C12Q 1/68 (2006.01) C12Q 1/70 (2006.01)

Patent

CA 2158883

Method and kits are provided for detecting one or more target nucleic acids. A first oligonucleotide having a 3' amino group and a second oligonucleotide having a 5' phosphate group are annealed to a contiguous complementary region of a target nucleic acid. Whenever the 3' terminal nucleotides of the first oligonucleotide and the 5' nucleotides of the second oligonucleotide are complementary to the opposing nucleotides on the target nucleic acid, a nucleic acid ligase ligates the first and second oligonucleotides via the formation of a phosphoramidate linkage. The presence of the target nucleic acid is determined by detection of the ligated first and second oligonucleotides.

éthodes et trousses pour la détection d'un ou de plusieurs acides nucléiques. Un premier oligonucléotide portant un groupe amine 3' et un second oligonucléotide portant un groupe phosphate 5' sont renaturés avec une région complémentaire contiguë de l'acide nucléique recherché. Lorsque les nucléotides de l'extrémité 3' du premier oligonucléotide et que ceux de l'extrémité 5' du second sont complémentaires avec les nucléotides situés vis-à-vis sur l'acide nucléique recherché, une ligase relie le premier et le second oligonucléotide en formant une liaison phosphoramidate. La présence de l'acide nucléique recherché est décelée par la mise en évidence du premier et du second oligonucléotide liés.

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