Patent ID: 7326575
Filing Date: 2008-02-05
Classification: C07H,C12Q,Y10S,Y10T

Abstract:
1. A method of in situ hybridization for detecting and specifically identifying a plurality of transcription sites in a cell, the method comprising: (a) treating a sample with ammonia-ethanol and sodium borohydride; (b) pressure cooking the sample; (c) assigning a different barcode to more than one different target sequence, wherein each barcode comprises at least one fluorochrome, and at least one barcode comprises at least two different spectrally distinguishable fluorochromes; (d) providing a probe set specific to the more than one different target sequences containing a hybridization probe specific for each target nucleic acid, each hybridization probe comprising a single nucleic acid molecule complementary to the target sequence, wherein each probe is labeled with a fluorochrome and the fluorochromes in each probe set collectively identify the barcode for the target sequence of that probe set; (e) incubating the tissue with a set of probes specific for each of the at least two different target sequences such that each hybridization probe simultaneously hybridizes to the target sequence to which each hybridization probe is complementary; and (f) detecting the fluorochromes on the probe set hybridized to RNA transcribed from each target sequence, if present, thereby separately detecting and specifically identifying the plurality of transcripts, wherein the detecting includes spectrally distinguishing the different fluorochromes, wherein the fluorochromes on a detected transcript constitute a barcode for the detected transcript.