Quantitation of mRNA by Competitive PCR Using Capillary Electrophoresis

The high sensitivity of the reverse transcriptase-polymerase chain reaction (RT-PCR) allows the amplification of low abundance mRNA transcripts, but the biomedical research and diagnostics community have increasing interest in the ability to accurately quantify the amounts of mRNA in cell samples. Although theoretically it should be possible to calculate the starting copy number from a consideration of the efficiency of the reverse transcription step of the reaction and the number of PCR cycles performed, it is widely recognized that this approach is flawed and can lead to highly inaccurate results (1 ).