Quantitative analysis of NQO1 gene expression by RT-PCR and CE-LIF.

ABSTRACT

A capillary electrophoresis-laser-induced fluorescence (CE-LIF) method to quantitate reverse transcription-polymerase chain reaction (RT-PCR) products of NAD(P)Hquinone acceptor oxidoreductase (NQO1) derived from whole blood after amplification with a reaction-specific internal standard is reported. The internal standard eliminates variability within the PCR (Hoffman-La Roche, Inc., Nutley, NJ, U.S.A.), while analysis by CE-LIF adds sensitivity and reduces variability associated with isotopic detection. Both the PCR and CE aspects of the assay are precise, with migration time precision of less than 1% and peak area ratio precisions of 9.8-15%. Future applications of this technique may include the analysis of gene therapy, oligonucleotides, and point mutations.