Rapid production of oligonucleotides

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/68 (2006.01)

Patent

CA 2567735

It has been previously disclosed that DNA segments can be made in massivel parallel chemical synthesis operations on a common substrate followed by release of the segments from the substrate and assembly of the segments into target DNA molecules. Here it is taught that if the DNA primary constructs are sufficiently long and properly designed, that the copy numbers of the primary constructs can be multiplied as needed by a PCR process using as a template regions at the ends of the primary constructs. The end regions, called flanking regions, can also be designed so that they may be cleaved easily from the amplification products. The target DNA can then be assembled from the cleaved fragments. Hundreds of thousands of oligonucleotides can be synthesized and assembled into many different individual genes by this process in a relatively quick and efficient process.

Il a été auparavant exposé qu'on peut fabriquer des segments d'ADN au cours d'opérations de synthèse chimique massivement en parallèle sur un substrat courant suivies de la libération des segments du substrat et de l'assemblage des segments en des molécules d'ADN cibles. Il est enseigné ici que si les constructions primaires d'ADN sont suffisamment longues et correctement conçues, les nombres de copies des constructions primaires peuvent être multipliés selon la nécessité par un procédé d'ACP utilisant comme matrice des régions aux extrémités des constructions primaires. Les régions des extrémités, appelées régions flanquantes, peuvent également être conçues de façon à ce qu'elles puissent être clivées facilement des produits d'amplification. L'ADN cible peut alors être assemblé à partir des fragments clivés. On peut synthétiser et assembler des centaines de milliers d'oligonucléotides en de nombreux gènes particuliers différents par ce procédé au cours d'un procédé relativement rapide et efficace.

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