Method for sequencing dna using a microfluidic device

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/68 (2006.01) B01L 3/00 (2006.01) B29C 59/16 (2006.01)

Patent

CA 2357775

The present invention describes a method for sequencing DNA in a device with microfluidics properties and a set of reagents for its use. This microfluidic device may be in form of a disc with radially extending microchannel structures (CD form) having an inner application area that may be common for one or more microchannel structures. By spinning the disc the liquid can be driven from an applicator area into reaction and/or detection are as closer to the periphery of the disc. Liquid transportation may thus be driven by centripetal force. The microfluidic device may also have other gemoetrical forms. Several methods can be used to determine the sequence of DNA according to the invention but the real time determination of released pyrophosphate using the luciferase luciferin reaction is preferred.

L'invention porte sur un procédé de séquencage d'ADN à l'aide d'un dispositif microfluidique et de son ensemble de réactifs, pouvant consister en un disque muni de micro-canaux radiaux (en forme de CD) et présentant une zone intérieure d'application pouvant être commune à l'un ou plusieurs des micro-canaux. Lorsque le disque tourne, le liquide est entraîné de la zone d'application vers le la zone de réaction/détection plus proche de la périphérie, la migration du liquide de faisant sous l'effet de la force centrifuge. Le susdit dispositif peut également présenter d'autres formes géométriques, et on peut dans le cadre de l'invention recourir à différentes méthodes pour déterminer une séquence d'ADN, la préférée consistant à déterminer en temps réel le pyrophosphate libéré, à l'aide de la réaction luciférase-luciférine.

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