Probe masking method of reducing background in an...

C - Chemistry – Metallurgy – 12 – P

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C12P 19/34 (2006.01) C07H 21/00 (2006.01) C12N 15/10 (2006.01) C12Q 1/68 (2006.01) C12Q 1/70 (2006.01)

Patent

CA 2223050

A method for reducing background caused by target-independent generation of amplification products, typically the products of a ligase chain reaction or a polymerase chain reaction, involves chemically "masking" or blocking the amplification probes or primers so that they cannot be extended or ligated until the occurence of a triggering event which can be delayed until the amplification reaction is begun. The probe masks take the form of complementary blocking oligonucleotides that are incapable of serving as template themselves and inhibit random tailing of the probe/primers. The blocking oligo masks are denatured from the probes during amplification and preferably are effectively eliminated from competing for probes in the amplification reaction

Procédé servant à limiter le signal d'arrière-plan provoqué par la génération, indépendante de la cible, de produits d'amplification, particulièrement, les produits d'une réaction de ligase ou de polymérase en chaîne. Ce procédé consiste à "masquer" ou à bloquer chimiquement les sondes ou les amorces d'amplification, de sorte qu'elles ne peuvent ni être allongées ni ligaturées jusqu'à l'apparition d'un évènement d'amorçage qui peut être retardé jusqu'au commencement de la réaction d'amplification. Ces masques de sonde prennent la forme d'oligonucléotides de blocage complémentaire incapables de servir de gabarits et inhibant l'extension homopolymérique au hasard des sondes et des amorces. Ces masques d'oligonucléotides de blocage apparaissent par dénaturation des sondes pendant l'amplification et, de préférence, sont éliminés, de manière à ne pas entrer en concurrence avec les sondes pendant la réaction d'amplification.

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