Improved procedure for the solid phase synthesis of...

C - Chemistry – Metallurgy – 07 – H

Patent

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C07H 21/00 (2006.01)

Patent

CA 2199246

An improved method of solid phase oligonucleotide synthesis is presented. The method results in lesser contamination of the final product with N-1, N-2, and other failure sequences. The method comprises employing a second capping step after initial loading and capping of the solid support and before addition of the second nucleoside. Further diminution in the N-1 oligonucleotide content can be realized when this second capping stepped is used in combination with loading the support to higher densities than have previously been deemed acceptable and by cleaving the full length oligonucleotide from the solid support using mildly basic conditions.

L'invention concerne un procédé amélioré de synthèse d'oligonucléotides en phase solide. Le procédé permet de réduire la contamination du produit final par des séquences N-1, N-2 et autres séquences tronquées. Le procédé consiste à utiliser une seconde étape de coiffage après le chargement et le coiffage initiaux du support solide et avant l'addition du second nucléoside. On peut réduire d'avantage la teneur en oligonucléotide N-1 lorsque l'on utilise cette seconde étape de coiffage en combinaison avec le chargement du support à des densités supérieures à celles considérées préalablement comme acceptables, et par clivage de l'oligonucléotide de longueur totale à partir du support solide dans des conditions légèrement basiques.

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