See discussion in the text for the equation used to quantify mRNA copies. Oncogene profiles from two T-cell lines. The quantity of DNA in each band in the oncogene profile was determined using ImageQuant software. Repeatability of quantification of c-myc mRNA levels in a sample. The use of synthetic RNA standards, the FluorKit DFA, and the FluorImager allows rapid, reproducible determination of oncogene mRNA expression.
Absolute quantification allows comparison of results between experiments and between laboratories.
Fluorescent labeling and detection of RT-PCR products provides sensitivity that is comparable to radioactive detection methods. The method can be applied to any system that requires measurement of mRNA levels. Direct analysis of gels on the FluorImager provides rapid, reproducible, quantitative results. Together the instrument, reagent system, and quantification software complement quantitative PCR by providing the accuracy, sensitivity, and dynamic range needed to assess mRNA levels in very small samples. Quantitative PCR Using Storage Phosphor Technology.