Methods and reagents for shortening incubation times in...

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/68 (2006.01) C40B 30/04 (2006.01)

Patent

CA 2747479

The methods and reagents described herein can be used to shorten incubation times in hybridization assays. As demonstrated in the examples, we have identified specific sulfonic acid polymers and hybridization conditions that lead to significantly shorter incubation times (e.g., signals after three hours that are comparable to signals that could traditionally only be obtained after overnight incubation). In some embodiments, shorter incubation times are achieved by adding the sulfonic acid polymer(s) during the hybridization process. Alternatively or additionally, in some embodiments, shorter incubation times are achieved via changes to the hybridizing conditions, e.g., by reducing the hybridization volume, increasing the salt concentration, and/or increasing the probe concentration (capture extender probe and/or label extender probe).

L'invention porte sur des procédés et des réactifs qui peuvent être utilisés pour raccourcir des temps d'incubation dans des dosages par hybridation. Comme démontré dans les exemples, nous avons identifié des polymères d'acide sulfonique spécifiques et des conditions d'hybridation qui conduisent à des temps d'incubation significativement plus courts (par exemple, signaux après trois heures qui sont comparables à des signaux qui ne pourraient être obtenus de manière classique qu'après une incubation pendant la nuit). Dans certains modes de réalisation, les temps d'incubation plus courts sont obtenus par l'addition du ou des polymères d'acide sulfonique durant le processus d'hybridation. En variante ou en plus, dans certains modes de réalisation, des temps d'incubation plus courts sont obtenus par des changements apportés aux conditions d'hybridation, par exemple par la réduction du volume d'hybridation, l'augmentation de la concentration en sel et/ou l'augmentation de la concentration en sonde (sonde d'extension de capture et/ou sonde d'extension de marquage).

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