USE OF A FLUORESCENT-PCR REACTION TO DETECT GENOMIC SEQUENCE COPY NUMBER AND TRANSCRIPTIONAL ABUNDANCE

We present a fluorescent-PCR-based technique to assay genomic sequence copy number and transcriptional abundance. This technique relies on t he ability to follow fluorescent PCR progressively in real time during the exponential phase of the reaction so that quantitative PCR is acc omplished. We demonstrated the ability of this technique to quantitate both known deletions and amplifications of loci that have been measur ed previously by other methods, and to measure transcriptional abundan ce. Using an efficient variant of the fluorescent-PCR technology, we c an monitor transcription semiquantitatively. The ability to detect all amplifications and deletions at any single copy locus by PCR makes th is the technique of choice to assay genomic sequence copy number anoma lies in birth defects and cancers. The ability to detect variations in transcript abundance enables this technique to fashion a time and tis sue analysis of transcription.