PROCESS FOR DETECTING LOW ABUNDANCE RNA IN INTACT CELLS

摘要

The present invention provides a process for detecting the presence of a target mRNA in an intact cell. The process includes the steps of (a) fixing and permeabilizing with a non-or weakly-crosslinking reagent the cell in the presence of a plurality of oligonucleotide probes, wherein each probe (i) contains about 15 to about 30 nucleotides, (ii) is labeled with a detectable marker, (iii) has a matched Tm of greater than about 60 DEG C, and (iv) specifically hybridizes to a different contiguous region of an open reading frame in the target mRNA with the provisos that each probe does not hybridize to itself, does not hybridize to any other probe and does not hybridize to a contiguous sequence of (A)n9(C)n9(G)n or (U)n in the target mRNA where n is an integer greater than 5; (b) removing unhybridized probes from the cell; and (c) measuring the detectable label in the cell.