Imaging individual mrna molecules using multiple singly...

Details Imaging individual mrna molecules using multiple singly... Imaging individual mrna molecules using multiple singly...

C - Chemistry, Metallurgy

12

Q

C12Q 1/68 (2006.01) C40B 30/04 (2006.01) C07H 21/00 (2006.01)

Patent

CA 2735197

A method for probing a target sequence of messenger ribonucleic acid molecules (mRNA's) in a fixed, permeabilized cell, said target sequence including at least 30 non- overlapping probe binding regions of 15-100 nucleotides, comprising immersing said cell in an excess of at least 30 nucleic acid hybridization probes, each singly labeled with the same fluorescent label and each containing a nucleic acid sequence that is complementary to a different probe binding region of said target sequence; washing said fixed cell to remove unbound probes; and detecting fluorescence from said probes.

La présente invention concerne un procédé pour rechercher une séquence cible de molécules d'acide ribonucléique messager (ARNm) dans une cellule fixée, perméabilisée, ladite séquence cible comprenant au moins 30 régions de liaison de sonde non chevauchantes de 15 à 100 nucléotides, comprenant l'immersion de ladite cellule dans un excès d'au moins 30 sondes d'hybridation d'acide nucléique, chacune ayant un marquage unique avec le même marqueur fluorescent et chacune contenant une séquence d'acide nucléique qui est complémentaire d'une région de liaison de sonde différente de ladite séquence cible ; le lavage de ladite cellule fixée pour éliminer les sondes non liées ; et la détection de la fluorescence desdites sondes.

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