Methods of determining the presence of double stranded...

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/68 (2006.01) C07H 21/04 (2006.01) C12N 15/10 (2006.01)

Patent

CA 2384100

Methods for determining the presence of double stranded nucleic acids in a sample are provided. As illustrated in the figure, nucleic acids present in a fluid sample are translocated through a nanopore, e.g. by application of an electric field to the fluid sample. The current amplitude through the nanopore is monitored during the translocation process and changes in the amplitude are related to the passage of single- or double-stranded molecules through the nanopore. The subject methods find use in a variety of applications in which the detection of the presence of double-stranded nucleic acids in a sample is desired, e.g. in hybridization assays, such as Northern blot assays, Southern blot assays, and array based hybridization assays.

L'invention concerne des méthodes permettant de déterminer la présence d'acides nucléiques double brin dans un échantillon. Comme illustré dans la figure 1A, les acides nucléiques présents dans un échantillon de fluide sont transloqués par un nanopore, par exemple par application d'un champ électrique sur l'échantillon de fluide. L'amplitude du courant dans le nanopore est surveillée pendant le processus de translocation et les changements d'amplitude sont liés au passage de molécules simple brin ou double brin par le nanopore. Ces méthodes sont utilisées dans diverses applications dans lesquelles on souhaite détecter la présence d'acides nucléiques double brin dans un échantillon, par exemple dans les tests d'hybridation, telles que les tests Northern blot, les tests Southern blot et les tests d'hybridation en série.

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