QUANTITATION OF RNA TRANSCRIPTS USING GENOMIC DNA AS THE INTERNAL AMPLIFICATION COMPETITOR

摘要

This invention describes a method for the quantitative monitoring of gene expression without either co-amplification of an added template or use of an endogenous constitutive transcript. The process involves a duplex amplification reaction in which a single set of primers is used to amplify both genomic DNA and expressed mRNA from the same gene sequence. These primers are targeted for sequences flanking the splice junction/intron sequences for the mRNA/DNA, as shown in the figure. Use of these primers with any suitable nucleic acid amplification technology will yield mRNA and DNA amplimers which are distinguishable by length and sequence heterogeneity. These amplimers will also be present in the final amplification reaction in ratios which are dependent upon the ratios of the expressed mRNA to the DNA in the sample, thus allowing thequantitation of mRNA in a sample.