Methods and means for amplifying nucleic acid

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/68 (2006.01)

Patent

CA 2474864

Methods of manipulation of nucleic acid, in particular amplification by means of the polymerase chain reaction (PCR), including use of oligonucleotides and combinations and kits comprising such oligonucleotides, also methods comprising use of nested PCR, allowing for improved results in methods wherein large numbers of nucleic acid fragments are manipulated by means of PCR and electrophoresis. Oligonucleotides are provided for use a size standards in electrophoresis, and internal controls allowing for calculation of relative amounts of material present. Improved results can be achieved in methods of profiling mRNA transcribed in a system under investigation.

La présente invention concerne des méthodes de manipulation d'acide nucléique, en particulier d'amplification au moyen de la réaction en chaîne de la polymérase (PCR), comprenant l'utilisation d'oligonucléotides ainsi que des combinaisons et des matériels comprenant lesdits oligonucléotides. L'invention concerne également des méthodes consistant à utiliser la PCR nichée qui permet d'obtenir des résultats améliorés dans des méthodes dans lesquelles des nombres importants de fragments d'acide nucléique sont manipulés au moyen de la PCR et de l'électrophorèse. L'invention concerne encore des oligonucléotides destinés à être utilisés en tant qu'étalons de masse moléculaire dans l'électrophorèse et des témoins internes permettant de calculer des quantités relatives de matière présente. De meilleurs résultats peuvent être obtenus dans des méthodes permettant de déterminer le profil de l'ARNm transcrit dans un système en cours d'examination.

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