Rapid direct sequence analysis of multi-exon genes

Details Rapid direct sequence analysis of multi-exon genes Rapid direct sequence analysis of multi-exon genes

C - Chemistry, Metallurgy

12

Q

C12Q 1/68 (2006.01) C07H 21/02 (2006.01) C07H 21/04 (2006.01) C12P 19/34 (2006.01) C12Q 1/00 (2006.01)

Patent

CA 2510891

Disclosed is a Single Condition Amplification/Internal Primer (SCAIP) sequencing method which allows for the rapid, accurate, and economical analysis of any large multi-exon gene. The method can be used to detect genomic mutations in any large multi-exon gene including the dystrophin gene. In some forms, the method can rely on amplification of a large number of exons at a single set of PCR temperatures with a first set of amplification primers followed by sequencing without optimization of individual amplicon conditions, using a second, internal set of sequencing primers. The SCAIP method provides for the identification and analysis of specific individual genomic mutations such as deletions, point mutations, frameshifts, or combinations thereof, in gene complexes with multiple exons/introns spanning large genomic regions.

La présente invention concerne une technique de séquençage par simple condition d'amplification/amorce interne (SCAIP) qui permet une analyse rapide exacte et économique de n'importe quel grand gène multi-exon. On peut utiliser cette technique pour détecter des mutations génomiques dans n'importe quel grand gène multi-exon, y compris le gène de la dystrophine. Dans certaines formes, cette technique repose sur l'amplification d'un grand nombre d'exons à un seul ensemble de température de PCR avec un premier ensemble d'amorces d'amplification suivi par le séquençage sans optimisation des conditions d'amplicon individuel, utilisant un deuxième ensemble d'amorce de séquençage. Cette technique SCAIP permet l'identification et l'analyse de mutations génomiques individuelles telles que des délétions, des mutations ponctuelles, des déphasages ou des combinaisons de ces mutations, dans des complexes géniques avec des exons/introns multiples portant sur de grandes régions génomiques.

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