QUANTITATIVE ANALYSIS OF GENE EXPRESSION

摘要

A method is provided for determining the level of expression of a gene of interest, or panel of genes of interest, in a biological sample. The method involves preparing a standard curve for each gene of interest by assaying a dilution series prepared from a total RNA in a reverse transcriptase-5'-exonuclease PCR amplification assay. A threshold cycle (Ct) for each member of the reverse transcribed-amplified is determined and plotted versus the log of the amount of total RNA in dilution series. The plot is used to determine an RNA equivalent from which the normalized RNA equivalent for the gene of interest in the sample is determined. The assay can be used to determine the effect of a treatment of the sample on the level of expression of a gene or interest or panel of genes of interest.