Gene synthesis by convergent assembly of oligonucleotide...

C - Chemistry – Metallurgy – 12 – P

Patent

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Details

C12P 19/34 (2006.01) C07H 21/00 (2006.01) C12M 1/00 (2006.01) C12N 15/10 (2006.01) C12Q 1/68 (2006.01)

Patent

CA 2723242

The invention provides a system and method for synthesizing polynucleotides by solid phase assembly oligonucleotide precursors, in accordance with the method, a polynucleotide is partitioned into an ordered set of subunits, wherein each subunit is assembled in a single reaction from a subset of oligonucleotide precursors that uniquely anneal together to produce the subunit. The subunits are then assembled to form the desired polynucleotide. An important feature of the invention is the selection of subunits that are free of undesired sequence elements, such as palindromes, repetitive sequences, and the like, which would result in more than one subunit product alter ligating a pool of oligonucleotide precursors.

L'invention concerne un système et un procédé destinés à synthétiser des polynucléotides par assemblage en phase solide de précurseurs oligonucléotides. Selon ledit procédé, un polynucléotide est divisé en un ensemble de sous-unités ordonnées, chaque sous-unité étant assemblée en une réaction unique à partir d'un sous-ensemble de précurseurs oligonucléotides qui recuisent uniquement ensemble pour produire la sous-unité. Les sous-unités sont ensuite assemblées pour former le polynucléotide souhaité. Une caractéristique importante de l'invention est que la sélection des sous-unités dépourvues d'éléments de séquence non désirés, tels que des palindromes, des séquences répétitives, et analogue, qui résulteraient en plus d'un produit de sous-unité modifiant la ligature d'un ensemble de précurseurs oligonucléotides.

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