Assaying nucleotides in solution using a fluorescent...

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/68 (2006.01) C12N 15/09 (2006.01)

Patent

CA 2295333

The invention provides a method for rapidly, economically and efficently sequencing and assaying nucleotides in a liquid medium using laser induced fluorescence of antisense probes, including PNA probes. Fluorescent intensity of the resulting medium is inversely proportional to the hybridization efficiency of the probes with respect to the target sequence. The method is particularly advantageous in not requiring separation of unhybridized probes and hybridization complexes prior to detection. The method can be used to identify accessible regions in folded nucleotide sequences, to determine the number of mismatched pairs in a hybridization complex, and to map genomes.

Cette technique permettant de séquencer et de doser, rapidement, économiquement et efficacement, des nucléotides dans un milieu liquide, utilise la fluorescence par laser de sondes antisens, notamment de sondes d'acide nucléique peptidique (PNA). L'intensité de la fluorescence du milieu résultant est inversement proportionnelle au taux de réussite d'hybridation des sondes relativement à la séquence cible. Cette technique s'avère des plus avantageuse dans la mesure où elle n'exige pas de séparation de sondes non hybridées et des complexes d'hybridation avant détection. On peut utiliser cette technique pour identifier des régions accessibles dans des séquences nucléotidiques pliées afin de déterminer le nombre de paires présentant un défaut de complémentarité dans un complexe d'hybridation et de cartographier des génomes.

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