Methods of synthesizing polynucleotides using thermostable...

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/68 (2006.01)

Patent

CA 2535922

Disclosed are methods of synthesizing a polynucleotide complementary to a target polynucleotide include steps of subjecting a non~-thermophilic cell comprising a thermostable polymerase to a temperature effective to disrupt the cell to form a reaction mixture, wherein the reaction mixture comprises the target polynucleotide and one or more primers that hybridize to a sequence of the target polynucleotide or to a sequence flanking the polynucleotide and incubating the reaction mixture under conditions whereby the polynucleotide is synthesized. Also disclosed are cell libraries and kits in accordance with the invention.

L'invention concerne des procédés permettant d'effectuer la synthèse d'un polynucléotide complémentaire d'un polynucléotide cible, qui consistent à soumettre une cellule non thermophile comprenant une polymérase thermostable à une température efficace permettant d'interrompre la cellule afin de former un mélange de réaction, ledit mélange de réaction comprenant le polynucléotide cible et une ou plusieurs amorce(s) qui s'hybrident avec une séquence flanquant ledit polynucléotide; et à mettre le mélange de réaction à incuber dans des conditions où la synthèse du polynucléotide est effectuée. L'invention concerne également des bibliothèques cellulaires et des kits selon l'invention.

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