High throughput genome sequencing on dna arrays

C - Chemistry – Metallurgy – 12 – Q

Patent

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Details

C12Q 1/68 (2006.01) C07H 21/04 (2006.01)

Patent

CA 2643700

The present invention is directed to methods and compositions for acquiring nucleotide sequence information of target sequences using adaptors interspersed in target polynucleotides. The sequence information can be new. e.g. sequencing unknown nucleic acids, re-sequencing, or genotyping. The invention preferably includes methods for inserting a plurality of adaptors at spaced locations within a target polynucleotide or a fragment of a polynucleotide. Such adaptors may serve as platforms for interrogating adjacent sequences using various sequencing chemistries, such as those that identify nucleotides by primer extension, probe ligation, and the like. Encompassed in the invention are methods and compositions for the insertion of known adaptor sequences into target sequences, such that there is an interruption of contiguous target sequence with the adaptors. By sequencing both "upstream" and "downstream" of the adaptors, identification of entire target sequences may be accomplished.

La présente invention concerne des méthodes et des compositions destinées à acquérir des informations de séquences nucléotidiques de séquences cibles au moyen d'adaptateurs intercalés dans des polynucléotides cibles. Les informations de séquences peuvent être nouvelles, notamment en matière de séquençage d'acides nucléiques inconnus, de re-séquençage ou de génotypage. L'invention concerne de préférence des méthodes destinées à insérer une pluralité d'adaptateurs au niveau d'emplacements espacés dans un polynucléotide cible ou un fragment d'un polynucléotide. Ces adaptateurs peuvent servir de plateformes pour interroger des séquences adjacentes au moyen de divers produits chimiques de séquençage, tels que ceux utilisés pour identifier des nucléotides par extension d'amorces, ligature de sondes et analogues. L'invention concerne également des méthodes et des compositions destinées à l'insertion de séquences d'adaptateurs connues dans des séquences cibles en vue d'une interruption d'une séquence cible contiguë avec les adaptateurs. Le séquençage "amont" et "aval" des adaptateurs permet de réaliser l'identification de séquences cibles dans leur intégralité.

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