Patent ID: 6534266
Filing Date: 2003-03-18
Classification: C12Q

Abstract:
An in situ hybridization method for detecting and specifically identifying transcription sites of a multiplicity of at least five different target sequences in a cell nucleus, the method comprising:assigning a different bar code to the at least five different target sequences, each of the target sequences comprising at least one predetermined subsequence, wherein each bar code comprises at least one fluorochrome, and at least one bar code comprises at least two different, spectrally distinguishable fluorochromes; providing a probe set specific for each of said at least five different target sequences, each probe set containing a hybridization probe specific for each predetermined subsequence, each hybridization probe consisting of a single nucleic acid molecule comprising a nucleotide sequence complementary to a predetermined subsequence in the target sequence, each probe being labeled with a fluorochrome, wherein the fluorochromes in each probe set collectively identify the bar code for the target sequence of that probe set; contacting the cell with a probe set specific for each of the at least five target sequences simultaneously such that each hybridization probe hybridizes to the predetermined subsequence to which each hybridization probe is complementary; and detecting fluorochromes on the probe set hybridized to RNA transcribed from each target sequence, if present, thereby separately detecting and specifically identifying transcription sites, wherein the detecting includes spectrally distinguishing the different fluorochromes; wherein the fluorochromes at a detected transcription site constitute a bar code for the detected transcription site.